Extended embryo culture up to 14 days | Medicina Reproductiva y Embriología Clínica

array:23 ["pii" => "S2340932022000068" "issn" => "23409320" "doi" => "10.1016/j.medre.2022.100118" "estado" => "S300" "fechaPublicacion" => "2022-09-01" "aid" => "100118" "copyright" => "Asociación para el Estudio de la Biología de la Reproducción y Sociedad Española de Fertilidad" "copyrightAnyo" => "2022" "documento" => "article" "crossmark" => 1 "subdocumento" => "rev" "cita" => "Medicina Reproductiva y Embriología Clínica. 2022;9:" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:1 [ "total" => 0 ] "itemSiguiente" => array:18 [ "pii" => "S2340932022000093" "issn" => "23409320" "doi" => "10.1016/j.medre.2022.100121" "estado" => "S300" "fechaPublicacion" => "2022-09-01" "aid" => "100121" "copyright" => "Asociación para el Estudio de la Biología de la Reproducción y Sociedad Española de Fertilidad" "documento" => "article" "crossmark" => 1 "subdocumento" => "rev" "cita" => "Medicina Reproductiva y Embriología Clínica. 2022;9:" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:1 [ "total" => 0 ] "en" => array:11 [ "idiomaDefecto" => true "cabecera" => "REVIEW" "titulo" => "Sperm DNA fragmentation and microfluidics: A new era in human sperm selection" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => array:2 [ 0 => "en" 1 => "es" ] "contieneResumen" => array:2 [ "en" => true "es" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:8 [ "identificador" => "f0005" "etiqueta" => "Fig. 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 4159 "Ancho" => 2953 "Tamanyo" => 618547 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "al0005" "detalle" => "Fig. " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "Microfluidic-based devices for sperm selection. A| First device based on sperm motility. It is consisted of 2-inlet and 2-outlet linked by channels. The exerted flow runs parallel to the passive sample current, which allows sorting of motile sperm in a separate chamber. B| Device based on sperm rheotaxis with 3 reservoirs and 4 channels. A flow is generated by hydrostatic pressure allowing the selection of sperm that swims against it. C| Microfluidic sorting device composed of two chambers connected to a micropump which generates a flow to select sperm according to rheotaxis behavior. D| Device consisting of a pool surrounded by U-shape channels. Different progesterone concentrations are added at inlet A while motile sperm is sorted at outlet B. E| Represents the CS-10 designed with an electric field which allows the transport of the motile sperm across a membrane. F| System that selects the motile sperm swimming fastest through a passive flow. G| Passive mechanism composed of a porous membrane capable of filtering sperm with better motility and fragmentation. H| The hydrodynamic behavior and swimming direction of the sperm is simulated in the SPARTAN for selection. It is made of 2 chambers connected by a channel composed of spaced micropillars. I| Based on boundary-following behavior, the selection in this device occurred by tracking sperm in a circular space of 500 microchannels. J| In this device, a patterned layer is placed forming a semi-circular structure while a covering layer is superimposed to establish inlet and outlet chambers to collect the highest quality sperm. K| The commercially available microchip is composed of 2 wells separated by a membrane with the capacity to filter sperm with better motility and fragmentation during a short incubation period. White sperm: non-motile. Blue sperm: motile. Green arrows indicate the direction of flow and red ones magnify the selected section. Adapted from A) <a class="elsevierStyleCrossRef" href="#bb0135">Cho et al., 2003</a> B) <a class="elsevierStyleCrossRef" href="#bb0535">Seo et al., 2007</a> C) <a class="elsevierStyleCrossRef" href="#bb0700">Zhang et al., 2015</a>a D) <a class="elsevierStyleCrossRef" href="#bb0705">Zhang et al., 2015b</a> E) <a class="elsevierStyleCrossRef" href="#bb0030">Ainsworth et al., 2005</a> F) <a class="elsevierStyleCrossRef" href="#bb0695">Zhang et al., 2011</a> G) <a class="elsevierStyleCrossRef" href="#bb0060">Asghar et al., 2014</a> H) <a class="elsevierStyleCrossRef" href="#bb0130">Chinnasamy et al., 2018</a> I) <a class="elsevierStyleCrossRef" href="#bb0400">Nosrati et al., 2014</a> J) <a class="elsevierStyleCrossRef" href="#bb0675">Xiao et al., 2021</a> K) <a class="elsevierStyleCrossRef" href="#bb0375">Meseguer et al., 2021</a>. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)" ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Maria Luisa Pardiñas, Angel Martin, David Ortega-Jaén, Jose María De los Santos, Thamara Viloria, Pilar Gamiz, Maria José De los Santos" "autores" => array:7 [ 0 => array:2 [ "nombre" => "Maria Luisa" "apellidos" => "Pardiñas" ] 1 => array:2 [ "nombre" => "Angel" "apellidos" => "Martin" ] 2 => array:2 [ "nombre" => "David" "apellidos" => "Ortega-Jaén" ] 3 => array:2 [ "nombre" => "Jose María" "apellidos" => "De los Santos" ] 4 => array:2 [ "nombre" => "Thamara" "apellidos" => "Viloria" ] 5 => array:2 [ "nombre" => "Pilar" "apellidos" => "Gamiz" ] 6 => array:2 [ "nombre" => "Maria José" "apellidos" => "De los Santos" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S2340932022000093?idApp=UINPBA00004N" "url" => "/23409320/0000000900000003/v1_202210200635/S2340932022000093/v1_202210200635/en/main.assets" ] "itemAnterior" => array:18 [ "pii" => "S234093202200007X" "issn" => "23409320" "doi" => "10.1016/j.medre.2022.100119" "estado" => "S300" "fechaPublicacion" => "2022-09-01" "aid" => "100119" "copyright" => "Asociación para el Estudio de la Biología de la Reproducción y Sociedad Española de Fertilidad" "documento" => "article" "crossmark" => 1 "subdocumento" => "rev" "cita" => "Medicina Reproductiva y Embriología Clínica. 2022;9:" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:1 [ "total" => 0 ] "es" => array:11 [ "idiomaDefecto" => true "cabecera" => "Revisión" "titulo" => "Aplicación de la inteligencia artificial en el laboratorio de reproducción asistida. Trabajo de revisión" "tienePdf" => "es" "tieneTextoCompleto" => "es" "tieneResumen" => array:2 [ 0 => "es" 1 => "en" ] "contieneResumen" => array:2 [ "es" => true "en" => true ] "contieneTextoCompleto" => array:1 [ "es" => true ] "contienePdf" => array:1 [ "es" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:8 [ "identificador" => "f0005" "etiqueta" => "Figura 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1056 "Ancho" => 1903 "Tamanyo" => 115689 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "al0005" "detalle" => "Figura " "rol" => "short" ] ] "descripcion" => array:1 [ "es" => "Esquema simplificado del proceso de laboratorio de reproducción asistida. El embriólogo realiza un análisis de semen del paciente masculino y evalúa los ovocitos de la paciente. En el caso de la técnica ICSI, se selecciona el mejor espermatozoide para inyectar en el ovocito. Durante el desarrollo embrionario se selecciona el embrión que se va a transferir y los embriones que se van a conservar mediante vitrificación, se descartan los embriones de desecho." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Paula Martín-Climent, Juan M. Moreno-García" "autores" => array:2 [ 0 => array:2 [ "nombre" => "Paula" "apellidos" => "Martín-Climent" ] 1 => array:2 [ "nombre" => "Juan M." "apellidos" => "Moreno-García" ] ] ] ] ] "idiomaDefecto" => "es" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S234093202200007X?idApp=UINPBA00004N" "url" => "/23409320/0000000900000003/v1_202210200635/S234093202200007X/v1_202210200635/es/main.assets" ] "en" => array:18 [ "idiomaDefecto" => true "cabecera" => "Review" "titulo" => "Extended embryo culture up to 14 days" "tieneTextoCompleto" => true "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "David Ortega-Jaen, Angel Martin, M.L. Pardiñas, Amparo Mifsud, Amparo Mercader, M.J. de los Santos" "autores" => array:6 [ 0 => array:3 [ "nombre" => "David" "apellidos" => "Ortega-Jaen" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "af0005" ] ] ] 1 => array:3 [ "nombre" => "Angel" "apellidos" => "Martin" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "af0010" ] ] ] 2 => array:3 [ "nombre" => "M.L." "apellidos" => "Pardiñas" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "af0010" ] ] ] 3 => array:3 [ "nombre" => "Amparo" "apellidos" => "Mifsud" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "af0005" ] ] ] 4 => array:3 [ "nombre" => "Amparo" "apellidos" => "Mercader" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "af0005" ] ] ] 5 => array:4 [ "nombre" => "M.J." "apellidos" => "de los Santos" "email" => array:1 [ 0 => "mariajose.delossantos@ivirma.com" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "af0005" ] 1 => array:2 [ "etiqueta" => "*" "identificador" => "cr0005" ] ] ] ] "afiliaciones" => array:2 [ 0 => array:3 [ "entidad" => "Laboratorio FIV, Instituto Valenciano de Infertilidad, IVI-RMA, Valencia, Spain" "etiqueta" => "a" "identificador" => "af0005" ] 1 => array:3 [ "entidad" => "Fundación IVI, Valencia, Spain" "etiqueta" => "b" "identificador" => "af0010" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cr0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author at: Laboratorio de FIV, Instituto Valenciano de Infertilidad, IVI-RMA, Pl. de la Policía Local, 3, Valencia 46015, Spain." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Cultivo embrionario extendido hasta 14 días" ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:8 [ "identificador" => "f0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 674 "Ancho" => 1778 "Tamanyo" => 119460 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "al0010" "detalle" => "Figure " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "Events of human implantation morphogenesis. The embryo formed by the ICM (inner cell mass) and the trophectoderm is shown first. On the one hand, from the ICM, the epiblast and hypoblast segregate, forming the bilaminar disc. In the epiblast, a cavity called the pro-amniotic cavity begins to appear, which will later give rise to the amniotic cavity, and the hypoblast extends to form the yolk sac. On the other hand, the trophectoderm differentiates into cytotrophoblast and syncytiotrophoblast." ] ] ] "textoCompleto" => "IntroductionHuman early embryonic development from long before pre-implantation to gastrulation has always been a very active but difficult field of developmental biology.It begins with a series of morphogenetic divisions and rearrangements leading to the formation of the blastocyst, constituted by the inner cell mass (ICM) and the trophectoderm, which floats freely between days 5 or 6 of development. Around the seventh day, this human embryo, now consisting of epiblast and primitive endoderm derived from the ICM and trophectoderm, must implant in the maternal uterus to survive (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>). Implantation of the blastocyst is a milestone in early mammalian embryonic development, as it is the moment at which a coordinated program of lineage diversification, cell fate specification and morphogenetic movements establishes the generation of extraembryonic tissues and the embryo proper, and determines the conditions for successful gestation and gastrulation (<a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al., 2016</a>).Errors in this process can lead to adverse pregnancy outcomes, including miscarriages and fetal defects, so understanding the mechanisms underlying this early development is of great importance not only for basic developmental biology but also for regenerative medicine (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>) and for resolving many cases of infertility and early pregnancy loss (<a class="elsevierStyleCrossRef" href="#bb0170">Lusine, 2020</a>).The first days of pre-implantation development have been extensively studied using methods pioneered in in vitro fertilization (IVF) (<a class="elsevierStyleCrossRef" href="#bb0075">Edwards et al., 1969</a>; <a class="elsevierStyleCrossRef" href="#bb0080">Edwards et al., 1970</a>). Most of our knowledge about human embryogenesis from pre-implantation to gastrulation derives from studies of existing anatomical and histological collections of human embryos (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>) and the majority of observations and experiments have been carried out using other mammals as models. For example, monkey embryos (<a class="elsevierStyleCrossRef" href="#bb0085">Enders et al., 1986</a>) as well as in vitro implantation systems in mice (<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al., 2014</a>) have been used as models for human embryo development.Recent advances in the in vitro embryogenesis, including the development of extended in vitro culture systems for mouse and primate embryos and the construction of stem cell-based embryo models, with the help of multi-omics studies and single cell-based imaging techniques, open new doors to study human embryonic development and have significantly improved our understanding of the characteristics and mechanisms of human embryos from pre-implantation to gastrulation (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>). However, although embryology presents a similar general program in all mammals, it should be noted that the extrapolation of data to humans is limited, because mammalian embryos show species-specific differences in post-implantation morphology (<a class="elsevierStyleCrossRef" href="#bb0240">Rossant, 2015</a>).Recently, in order to explain what happens in the early stages after human embryo implantation, the groups of <a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al. (2016)</a> and <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al. (2016)</a> have adapted a protocol for in vitro culture of mouse embryos, allowing human embryos to develop up to the maximum number of days legally allowed.In the present work we aim to review aspects related culture conditions, morphogenesis events, potential applications of long-term in vitro culture of human embryos as well as some ethical issues surrounding this topic.Materials and methodsIn the following study, an electronic bibliographic review of several papers and publications obtained from different databases, including Pubmed and the National Center for Biotechnology Information (NCBI), has been carried out. In addition, the academic search engine "Google Scholar" and different websites, such as ThermoFisher Scientific, have been used for some components of the culture medium. The research of information took place between the end of 2021 and the beginning of 2022, and the main materials used were papers in English or Spanish from the last five years. The keywords used for the research of information were: human embryo development, in vitro embryo development, pre-implantation, in vitro embryo culture, and 14-day rule.Results and discussionWhat is extended embryo culture up to 14 days and what are its conditions?This extended culture consists of culturing fresh or thawed embryos that are at day 5 or 6 of development up to the maximum of 14 days allowed by law. But how has it been possible to culture human embryos in vitro up to this limit?Before answering this question, it should be noted that, although long-term in vitro culture of mouse and primate embryos is promising, the current conditions need to be optimised in terms of reproducibility and efficiency. High-quality embryos and optimised cell culture environments (culture medium, supplements, and gas regulation mechanism) are essential for stable and efficient protocols for long-term mammalian embryo culture. Furthermore, in current long-term cultures, extra-embryonic tissues tend to grow vigorously with embryo development, hindering the development of the inner embryo by absorbing nutrients from the culture medium. In this sense, strategies to equilibrate extra-embryonic and embryonic growth may also be a good starting point to achieve long-term embryo culture, as human embryo development is highly dependent on the simultaneous growth of extra-embryonic tissues that provide essential signals to help extend embryo development to certain stages (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>).Meanwhile, the group of <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al. (2016)</a> established a culture system that allowed human embryos to develop in vitro through post-implantation stages. This was based on a system for culturing mouse embryos through implantation (<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al., 2014</a>).This procedure basically consists of thawing the embryos (if vitrified), removing the zona pellucida with tyrodes acid, and placing them in culture until day 14 (<a class="elsevierStyleCrossRef" href="#f0005">figure 1</a>). Interestingly, as in the sequential approach used in IVF to culture embryos from day 3 to blastocyst culture to provide the embryos with a specific nutrient environment, long-term culture requires a gradual replacement of culture media containing decreasing amounts of serum. The first 48 hours of culture that is from day 5-6 to day 7-8, embryos are cultured using first culture medium (IVC1). On day 9-10, half of the culture medium is replaced by another medium (IVC2) and renewed daily until day 14 by removing half of the medium and adding the other half. More detailed information can be found in <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al. (2016)</a> and in <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al. (2016)</a>.<elsevierMultimedia ident="f0005"></elsevierMultimedia>The conditions and components of the extended culture medium are explained belowLike conventional embryo culture, long-term culture has, with some particularities, similar culture conditions requirements to conventional embryo culture in IVF.TemperatureWith regard to the optimal culture temperature, it depends on the body temperature of the animal from which the cells are collected and the regional temperature variation in that animal. Some studies have shown that the temperature within the female reproductive tract can be lower than 37°C, indicating that a lower temperature may be more beneficial for embryo development (<a class="elsevierStyleCrossRef" href="#bb0015">Baak et al., 2019</a>). Indeed successful fertilization, blastocyst formation rates and pregnancies can be achieved at 36°C (<a class="elsevierStyleCrossRef" href="#bb0125">Hong et al., 2014</a>).Currently, the temperature used in the long-term culture is also 37°C (<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al., 2014</a>; <a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al., 2019</a>; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0260">Shahbazi et al., 2020</a>; <a class="elsevierStyleCrossRef" href="#bb0340">Weimar et al., 2013</a>; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al., 2016</a>).Gas environmentCarbon dioxide (CO2) and oxygen (O2) levels inside the culture incubator must also be controlled. CO2 is essential to maintain the right pH in the culture medium, which ranges between 7.2-7.4, and allow proper embryo development (<a class="elsevierStyleCrossRef" href="#bb0225">Reimundo et al., 2021</a>; <a class="elsevierStyleCrossRef" href="#bb0230">Rendón Abad, 2018</a>). As in IVF, 5% CO2 concentration is commonly used for embryo culture up to day 14 (<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al., 2014</a>; <a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al., 2019</a>; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0260">Shahbazi et al., 2020</a>; <a class="elsevierStyleCrossRef" href="#bb0340">Weimar et al., 2013</a>; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al., 2016</a>).Regarding O2 concentration, the trend in IVF clinics over the past years has been to culture pre-implantation human embryos in hypoxic conditions (5% O2) (<a class="elsevierStyleCrossRef" href="#bb0060">Christianson et al., 2014</a>). Low O2 tension improves both embryo development and IVF outcomes (<a class="elsevierStyleCrossRef" href="#bb0065">De los Santos et al., 2013</a>; <a class="elsevierStyleCrossRef" href="#bb0320">van Montfoort et al., 2020</a>). When data from the meta-analyses were stratified for duration of culture under low O2, it was demonstrated that even though embryo quality was lower at cleavage stage (<a class="elsevierStyleCrossRef" href="#bb0065">De los Santos et al., 2013</a>), pregnancy or live birth rates were only higher when the blastocyst stage (<a class="elsevierStyleCrossRef" href="#bb0190">Nastri et al., 2016</a>).As the embryo is more vulnerable to oxygen in the pre-compaction stage as compared to the post-compaction stage, low O2 is recommended from the initial stages of development (<a class="elsevierStyleCrossRef" href="#bb0325">Wale and Gardner, 2010</a>; <a class="elsevierStyleCrossRef" href="#bb0165">Kirkegaard et al., 2013</a>). However, it has been observed that beyond day 7 of development, embryos may benefit from higher O2 concentrations (21%), possibly due to the increased size of the embryo, which may decrease the O2 pressure in the embryonic nucleus. Hypoxia clearly has a complex influence on embryonic development and its effects need to be better understood in the intrauterine environment to determine exactly how it affects embryogenesis (<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>). With regard to the concentration of this gas, although some authors have used hypoxia conditions (<a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al., 2019</a>), most of them have done the extended culture until day 14 with an O2 concentration of 21% (<a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0260">Shahbazi et al., 2020</a>; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al., 2016</a>).Culture media compositionThe composition of the long-term culture medium is slightly modified compared to the embryo culture media widely used in IVF laboratories such as G-2TM (Vitrolife, Sweden) or Sydney IVF Blastocyst Medium (Cook Medical, United States) as they do not contain hormones, nor serum from animal sources.Indeed, both IVC1 and IVC2, are made up of the same components such as advanced DMEM/F12, ITS-X, L-glutamine, β-oestradiol, progesterone, penicillin/streptomycin, and N-acetyl-L-cysteine. Whereas IVC1 is supplemented with 20% heat-inactivated FBS, IVC2 by 30% Knockout Serum Replacement.Advanced DMEM/F12 (ThermoFisher Scientific, United States) which used as basal medium, is composed, like the conventional IVF media mentioned above, of amino acids, inorganic salts, vitamins, proteins, and glucose. However, it has certain peculiarities. It does not contain human serum albumin (HSA), but instead includes other proteins such as AlbuMAX® II (Gibco, United States), human transferrin and recombinant full-chain insulin. Furthermore, it contains additional components such as fatty acids like linoleic acid and linolenic acid, reducing agents like glutathione, monosodium, and other components like putrescine or thymidine (all components and concentrations are specified in <a class="elsevierStyleCrossRef" href="#t0005">Table 1</a>) (<a class="elsevierStyleCrossRef" href="#bb0300">ThermoFisher Scientific, 2020a</a>). Despite this, the in vitro function of some of these components on the embryo has not been completely described.<elsevierMultimedia ident="t0005"></elsevierMultimedia>On the other hand ITS-X, which is composed of: (1) insulin, which promotes glucose and amino acid uptake, lipogenesis, intracellular transport, and protein and nucleic acid synthesis; (2) transferrin, which acts as an iron carrier and can also help reduce toxic levels of oxygen radicals and peroxide; (3) selenium, as sodium selenite, which acts as a cofactor for glutathione peroxidase and other proteins that is used as an antioxidant in the medium; and (4) ethanolamine, a precursor of phosphoglycerides that are essential for the structure of the plasma membrane and cellular organelles (Concentrations are specified in <a class="elsevierStyleCrossRef" href="#t0005">Table 1</a>). All these components have been shown to reduce the need for serum supplementation of conventional cell culture media and to promote optimal performance of serum-free media (<a class="elsevierStyleCrossRef" href="#bb0290">ThermoFisher Scientific, 2014b</a>). This is the case for the culture media up to day 14, IVC1 and IVC2 respectively, where IVC1 contains only 20% inactivated FBS and IVC2 contains Knockout Serum Replacement, which replaces FBS. Possible reasons for using reduced serum reduced serum are unknown exact composition, batch-to-batch variability, unintended interaction with culture substances, potentially leading to unexpected or undesired results, experimental variability, and limited reproducibility (<a class="elsevierStyleCrossRef" href="#bb0315">van der Valk et al., 2018</a>). In addition, with Knockout Serum Replacement, many of the disadvantages of using FBS in culture can be mitigated (<a class="elsevierStyleCrossRef" href="#bb0300">ThermoFisher Scientific, 2020a</a>).As discussed, both conventional and long-term culture media contain amino acids in their composition. These molecules have numerous functions, and their importance is attributed to acting as precursors for protein and nucleotide biosynthesis, substrates for energy metabolism, osmolytes, antioxidant, regulators, buffers, and chelators. From studies in mice, non-essential amino acids stimulate the rate of cleavage, blastocyst formation and hatching. Essential amino acids stimulate cleavage rates after the eight-cell stage and increase the development of the inner cell mass in the blastocyst (<a class="elsevierStyleCrossRef" href="#bb0280">Sunde et al., 2016</a>).As in IVF culture media, L-glutamine is not present, instead GlutaMAXTM (ThermoFisher Scientific, United Stated) is added as an alternative. This supplement consists of the dipeptide L-alanyl-L-glutamine, which is more stable in aqueous solutions, making it suitable for suspension and adherent cultures of mammalian cells, such as embryos cultured up to day 14. Moreover, compared to L-glutamine, GlutaMAX minimises toxic ammonia accumulation, improves cell viability and growth by having lower ammonia levels, and remains stable over a wide temperature range because it does not spontaneously degrade (<a class="elsevierStyleCrossRef" href="#bb0305">ThermoFisher Scientific, 2020b</a>). It should be noted that, although this supplement as such is not added to conventional culture media used in IVF laboratories, they contain stabilised glutamine in the form of L-alanine-L-glutamine.Another peculiarity of the basal medium of the long-term culture is that it contains a compound called AlbuMAX® II. Conventional culture medium contains HSA with associated lipids. However, this supplement consists of lipid-rich bovine serum albumin (BSA), and like ITS-X, is effective in reducing or replacing the requirement for serum supplementation in culture media. In addition, the chromatographic separation process contributes to the preservation of the natural lipids associated with the purified albumin, resulting in excellent growth stimulation characteristics (<a class="elsevierStyleCrossRef" href="#bb0295">ThermoFisher Scientific, 2014c</a>).Additionally, to these protein-associated lipids, the long-term culture medium contains other lipids, specifically fatty acids, which are not added normally in conventional IVF media such as those mentioned above. It is known that embryos need lipids at very early stages. Most of the ATP for pre-implantation development comes from fatty acids. The proportion of different types of lipids plays an important role. An increase in unsaturated fatty acids can compensate for the negative effects of saturated fatty acids. Polyunsaturated fatty acids (linoleic acid and linolenic acid) have a beneficial role in embryonic cells signalling and may influence gene expression in lipid metabolism, oxidation and fatty acid synthesis as seen in animal models (<a class="elsevierStyleCrossRef" href="#bb0210">Pawar and Jump, 2003</a>). The impact of trans-unsaturated fatty acids on embryonic development is unknown. However, it is thought that their effect on polyunsaturated fatty acid metabolism could affect embryonic development (<a class="elsevierStyleCrossRef" href="#bb0095">Fürstova et al., 2008</a>). This means that lipid balance may be important for pre-implantation embryo development and implantation (<a class="elsevierStyleCrossRef" href="#bb0355">Zander-Fox et al., 2021</a>).The basal medium also contains, although at low concentration, polyamines such as putrescine. These play a key role in cell proliferation, growth, and differentiation. They are involved in the regulation of DNA and protein synthesis, proliferation and differentiation (<a class="elsevierStyleCrossRef" href="#bb0160">Igarashi and Kashiwagi, 2000</a>). They act as antioxidants protecting DNA, proteins and lipids from oxidative damage (<a class="elsevierStyleCrossRef" href="#bb0055">Chattopadhyay et al., 2003</a>). There are evidences that these molecules regulate early embryogenesis, placental trophoblastic growth, implantation and embryonic development (<a class="elsevierStyleCrossRef" href="#bb0110">Halloran et al., 2021</a>; <a class="elsevierStyleCrossRef" href="#bb0350">Wu and Morris Jr, 1998</a>). In addition, supplementation of polyamines such as putrescine has been shown to be involved in embryo survival, ultimately resulting in improved fertility (<a class="elsevierStyleCrossRef" href="#bb0145">Hussain et al., 2017</a>).Another component present in the culture media is vitamins. In contrast to conventional media, the culture up to day 14 contains an abundance of vitamins B and C. Although the role of some of them in embryonic development is known, the role of others is unknown. In hamsters, vitamins B have been shown to promote embryonic development in vitro; however, they may be inhibitory in early mouse embryos. Vitamins B2 (riboflavin), 6 (pyridoxine), 9 (folic acid), 12 (cobalamin) are known to be necessary for homocysteine recycling (<a class="elsevierStyleCrossRef" href="#bb0185">Ménézo et al., 2013</a>). Vitamin C (ascorbic acid) acts as an antioxidant by reducing oxidative stress-induced embryo toxicity, and in mice it also improves blastocyst development rates (<a class="elsevierStyleCrossRef" href="#bb0330">Wang et al., 2002</a>).In addition to the mentioned molecules that act as antioxidants (amino acids, proteins, vitamins), the basal culture medium of the extended culture contains N-acetyl-L-cysteine and reducing agents such as glutathione, monosodium which are not present in conventional IVF media.As mentioned before, the culture medium up to day 14 also contains hormones which are not present in the routine culture media used in IVF laboratories. There is a wealth of evidence that growth factors and hormones in the reproductive tract and in the pre-embryo may be potential regulators of embryonic development. Their benefits are thought to include decreased apoptosis, increased cell proliferation, and consequently improved progression to blastocyst. The need for their presence and usefulness in culture media is unclear due to the unknown long-term effects they may have on pre-embryos. However, culture media containing different growth factors and hormones can be found on the market (<a class="elsevierStyleCrossRef" href="#bb0230">Rendón Abad, 2018</a>). Long-term culture medium, in its initial protocol and unlike conventional IVF media, contains progesterone and β-oestradiol.Extended culture medium will also include other supplements such as antibiotics (e.g. penicillin and streptomycin). These are added to the medium to prevent or reduce likelihood of contamination. Although gentamicin has been found to be the antibiotic used par excellence in culture media as a prophylactic measure, it should be noted that the absence of antibiotics is beneficial for the development of pre-embryos as they develop faster (<a class="elsevierStyleCrossRef" href="#bb0230">Rendón Abad, 2018</a>).What structures can we see with the culture up to day 14?In vitro culture systems up to day 14 of embryo development have made it possible to document the key events of normal human embryogenesis from pre-implantation to gastrulation. By comparing embryos developing in vitro with the Carnegie series of human embryos in vivo (<a class="elsevierStyleCrossRef" href="#bb0120">Hertig et al., 1956</a>), the main morphogenetic events of human implantation morphogenesis were discovered (<a class="elsevierStyleCrossRef" href="#f0010">figure 2</a>). These include:<ul class="elsevierStyleList" id="l0005"><li class="elsevierStyleListItem" id="li0005">1.Segregation of the epiblast and hypoblast from the inner cell mass of the blastocyst and morphogenetic movements to form the bilaminar disc. While the epiblast is represented by increased levels of OCT4 and Nanog (pluripotency transcription factors), the hypoblast is determined by increased levels of GATA6 and Sox17 (hypoblast-specific transcription factors).</li><li class="elsevierStyleListItem" id="li0010">2.Pro-amniotic cavity formation within pluripotent epiblast cells of the embryonic lineage (via apically located atypical PKC, leading to apical polarity in epiblast cells and lumen formation) and amniotic epithelium.</li><li class="elsevierStyleListItem" id="li0015">3.Formation of the embryonic disc from the epiblast (embryo proper).</li><li class="elsevierStyleListItem" id="li0020">4.Formation of the primary yolk sac within the hypoblast.</li><li class="elsevierStyleListItem" id="li0025">5.Differentiation of the trophoblast to produce its two characteristic cell types: the mononucleated cytotrophoblast and the multinucleated polyploid syncytiotrophoblast.</li></ul><elsevierMultimedia ident="f0010"></elsevierMultimedia>Despite not all embryos will be able to undergo the former morphogenic event, the system brings in vitro cultured embryos to the point of gastrulation (<a class="elsevierStyleCrossRef" href="#bb0250">Shahbazi and Zernicka-Goetz, 2018</a>). Interestingly, all of them can take place in vitro in the absence of maternal tissues, indicating that human embryos have a previously underestimated potential for self-organization (<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al., 2016</a>).Although this system may not be able to fully recapitulate all aspects of human embryogenesis in vivo, it has revealed that even in the absence of endometrial cells, human blastocysts have remarkable capacity for self-organization that was previously unknown (<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>).Stem cells and cultureIn recent years, the cultivation of stem cell-based embryonic models to unravel the mysteries of early embryonic development in humans has gained ground.The stem cell-based embryo model is a simplified model that attempts to mimic the characteristics of a natural embryo. The knowledge gained from their experimentation opens a new way to model embryogenesis, cell lineage differentiation, tissue morphogenesis, and organogenesis in mammalian development (<a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam, 2021</a>). Moreover, it can improve the understanding of the mechanisms of embryo development from blastocyst stage to gastrulation, as it is more scalable, versatile, accessible than natural embryos, and exhibits similar self-organization as natural embryos during in vitro culture (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>).There are two types of stem cell-based models (<a class="elsevierStyleCrossRef" href="#f0015">figure 3</a>). On the one hand, non-integrated models use a single stem cell type (ESC) and are limited to the formation of specific parts of the embryo such as amniotic sac structures and shed light on selected morphogenetic events such as gastrulation or developmental processes such as germ layer differentiation. This type of model includes amniotic sac models (<a class="elsevierStyleCrossRef" href="#bb0370">Zheng et al., 2019</a>) and the elongated gastruloid resembling the posterior elongation body axis (<a class="elsevierStyleCrossRef" href="#bb0020">Beccari et al., 2018</a>). On the other hand, integrated models are formed from several stem cell types (ESC, XENC, TSC) and contain all parts of the conceptus, that is, embryonic and extra-embryonic tissues, giving rise to the blastoid, a blastocyst-like structure formed by the aggregate of ESC and TSC, where TSC encloses ESC in a cavity (<a class="elsevierStyleCrossRef" href="#bb0235">Rivron et al., 2018</a>) and the embryoid, an embryo-like structure made up of various types of stem cells but not capable of developing into a functional human embryo (<a class="elsevierStyleCrossRef" href="#bb0270">Sozen et al., 2018</a>). Both types of models can be achieved by means of 2D micropatterning techniques, microfluidic devices, or 3D aggregation methods, exhibiting a self-organization similar to that of natural embryos during in vitro culture. It should be noted that most of this type of work has now been carried out in animal models such as the mouse (<a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam, 2021</a>).<elsevierMultimedia ident="f0015"></elsevierMultimedia>Limited knowledge about the mechanisms regulating cell fate determination during early human embryogenesis hampers the creation of stem cell-based embryo models. This could only be remedied by further research with human embryos per se, non-human primate embryos in vitro and embryo models (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>). And, although these culture systems could reproduce the events of early mammalian embryogenesis in vivo, a critical prerequisite of experimental models is that they must accurately reconstruct the events that occur in vivo, particularly in the early stages when the basic body plan is established and when the in vivo embryo is most inaccessible for experimentation (<a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam, 2021</a>). By collecting more data on this, extended ex vivo phenotype, new markers of embryo competence and sustained implantation could be identified to enhance IVF outcomes.Potential applications in embryologyThe contributions of post-implantation embryo research, apart from allowing to observe in vitro the events and structures mentioned above, could include advancing the understanding of gene function during embryogenesis; a scientific basis for the prevention of early pregnancy loss, birth defects and teratogenesis; and an understanding of how the widespread epigenetic programming that occurs during this stage of development might affect disease progression in later life (<a class="elsevierStyleCrossRef" href="#bb0215">Pera, 2017</a>).Advances in in vitro culture systems have allowed the modelling of chromosomal instability and the investigation of how mosaicism and chromosomal abnormalities can affect development in the pre-implantation period.Before post-implantation in vitro culture, the capacity of mosaic blastocysts had only been studied in mice (<a class="elsevierStyleCrossRef" href="#bb0035">Bolton et al., 2016</a>) or through clinical outcome data (<a class="elsevierStyleCrossRef" href="#bb0100">Greco et al., 2015</a>) with no blinded studies evaluating the IVF outcome of the reported mosaicism. Without knowing the implications of mosaic embryos in IVF transfers, it is difficult to determine whether to discard them and waste a potentially viable embryo or transfer a potentially harmful one. <a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al. (2019)</a>, using NGS and extended culture to day 12 of development, are believed to be the first to investigate the fate of chromosomal abnormalities and mosaicism. On the one hand, they showed that many mosaics in the day 5 or 6 blastocyst biopsy were no longer mosaic at 8 or 12 days, others were still mosaic, and others had become uniformly aneuploid. In general, when mosaic blastocysts had more abnormal cells, they were more likely to be non-viable at 12 days. On the other hand, they also showed that NGS can have an 18% misdiagnosis rate for mosaicism, so improvements and further studies are needed before clinical application. However, it shows the possible developments of these advances in extended culture (<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson, 2020</a>).Consensus on the clinical management of embryonic mosaicism is still under debate. In vitro performance of specific mosaics are needed to understand their developmental behaviour and improve diagnosis. Furthermore, assessing chromosomal instability during these hitherto hidden developmental stages may provide valuable information on the predictive value of reporting mosaicism in clinical practice, ultimately improving the embryo selection process and clinical outcomes.14-day limitThe 14-day limit for in vitro human embryo culture dates back to the 1970s, more specifically to 1978, when the ability to develop embryos in vitro for research and assisted reproduction was demonstrated (<a class="elsevierStyleCrossRef" href="#bb0275">Steptoe and Edwards, 1978</a>). But it was not until 1984 that it was first established in the Warnock Report in the UK (<a class="elsevierStyleCrossRef" href="#bb0335">Warnock, 1984</a>), and subsequently enshrined in the <a class="elsevierStyleCrossRefs" href="#bb0130">Human Fertilization and Embryology (HFE) Acts of, 1990, 2008</a>. It is now one of the most internationally accepted standards in reproductive medicine (<a class="elsevierStyleCrossRef" href="#bb0010">Appleby and Bredenoord, 2018</a>).The Warnock Report (1984) elicited different responses from the public and committee members as there was a wide range of opinions on the moral status of human embryos. However, most of the committee agreed that such research should be allowed under licence. To allay public fears it was necessary to set a clear limit, which was 14 days, that could be justified on the basis of this moral status of the embryo (<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger, 2021</a>).One of the reasons for setting the maximum at 14 days was that this point in development was prior to the formation of the primitive line, which appears on day 15 of human embryonic development. This occurs at the onset of gastrulation, a period when the three germ layers are formed and the axes and body form are established, marking the point of individualisation (<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson, 2020</a>). The committee's view was that from this point onwards the embryo could be considered as a defined individual and a potential person, an entity with rights, such as the right to life, since from this point onwards twins cannot be formed. Furthermore, he pointed out that this boundary was prior to the formation of the central nervous system, around day 22 of development, which would exclude the possibility of the embryo experiencing pain (<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger, 2021</a>). Other reasons were the substantial loss of embryos from fertilization to this point and that until the implantation process is not complete, the embryo has no potential for further development (<a class="elsevierStyleCrossRef" href="#bb0215">Pera, 2017</a>).However, given that scientific progress has brought us to a point where viable pregnancies can be obtained from day 7 blastocyst transfer, could be reasonable to question whether 14 days is too restrictive from a research point of view (<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson, 2020</a>). Furthermore, as discussed before, recent studies have demonstrated the successful growth of human embryos up to developmental stages close to this limit (<a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>). Other research has reported the generation of embryo-like structures from mouse and human pluripotent stem cells that mimic the gastrulating embryo in form and cellular content (<a class="elsevierStyleCrossRef" href="#bb0090">Etoc et al., 2016</a>; <a class="elsevierStyleCrossRef" href="#bb0115">Harrison et al., 2017</a>; <a class="elsevierStyleCrossRef" href="#bb0265">Shao et al., 2017</a>; <a class="elsevierStyleCrossRef" href="#bb0310">van den Brink et al., 2014</a>). And recently, successful in vitro culture of Cynomolgus macaque embryos beyond the gastrula stage and up to day 20 of development has been reported (<a class="elsevierStyleCrossRef" href="#bb0195">Niu et al., 2019</a>). This may fill the knowledge gap on human embryogenesis beyond day 14, and it is even suggested that such a culture period should also be feasible for human embryos (<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson, 2020</a>).All these advances have led to a re-examination of the current restrictions on human embryo culture (<a class="elsevierStyleCrossRef" href="#bb0150">Hyun et al., 2016</a>), with various positions or sides emerging. There are those in favour of such an extension (<a class="elsevierStyleCrossRef" href="#bb0155">Hyun et al., 2021</a>; <a class="elsevierStyleCrossRef" href="#bb0175">McCully, 2021</a>); others who think that it should not be extended and should remain as it currently stands (<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger, 2021</a>; <a class="elsevierStyleCrossRef" href="#bb0105">Green et al., 2021</a>; <a class="elsevierStyleCrossRef" href="#bb0180">McLaren, 1984</a>); and even the most extreme who argue that human embryos should not be experimented on. Making changes to existing legislation is a complex process and there is a risk of creating new restrictions that could have unintended consequences for future research (<a class="elsevierStyleCrossRef" href="#bb0215">Pera, 2017</a>).So, should the 14-day limit be extended?The period between days 14 and 28 of human embryonic development is called the "black box" (<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al., 2017</a>). Until now, embryonic development in this period has been difficult to study and understand from a scientific and regulatory point of view. However, the <a class="elsevierStyleCrossRef" href="#bb0200">Nuffield Council on Bioethics (2017)</a> argues that any attempt to extend research beyond day 14 would have to result in significant advances in science. Furthermore, it should examine the committee's reasoning and justify the extension on this basis. The two key considerations are maintaining public confidence and providing a moral justification for embryo research. The committee provided two reasons for morally justifying early embryo research: the utilitarian argument, based on the benefits of such research, and its argument based on the moral status of early embryos (<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger, 2021</a>).Due to the current benefits of embryo research and the potential future benefits, supporters of extending the 14-day rule are calling for its revision. Extending this period to 28 days would allow to study the developmental processes of gastrulation, when the first primitive tissues are formed. In addition, it would allow learning about the developing nervous system without the risk of neural connections and a better understanding of early organ development (<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al., 2017</a>). It would also be possible to advance our understanding of cell fate decisions during early embryonic development (<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al., 2016</a>). The safety and success of current IVF procedures could be further improved by understanding the nature of some birth defects and predicting which embryos are most likely to result in a successful pregnancy (<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al., 2017</a>). Finally, more could be learned about the physiology of pregnancy beyond day 14, including the processes surrounding implantation, a greater understanding of the causes of embryonic defects and why events such as miscarriages occur (<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al., 2017</a>). <a class="elsevierStyleCrossRef" href="#bb0045">Castelyn (2020)</a> offers several reasons why a re-evaluation of the 14-day rule may be justified.Sophia McCully takes this side, listing several potential advantages to extending the limit to day 28 of development (<a class="elsevierStyleCrossRef" href="#bb0175">McCully, 2021</a>). She focuses mainly on the utilitarian argument to justify her idea, suggesting that it will help reduce miscarriage rates, improve IVF outcomes, and test the safety of new techniques such as gene editing. And, while acknowledging that the current limit has generated public confidence, she argues that concerns about a slippery slope are unjustified because the regulations are strict, making it illegal to slide down such a slope. Other authors advocate a more conservative extension of about 2-3 days, and that only after broad public consultation (<a class="elsevierStyleCrossRef" href="#bb0155">Hyun et al., 2021</a>).However, although it has important potential scientific and medical benefits, it is unclear whether extending the 14-day limit in human embryo culture is the best way to achieve them or not, as the possibility of prolonged normal development of human embryos in vitro during and beyond the primitive line stage is unknown. In mice, pre-implantation embryo culture has so far not generated structures that resemble post-implantation embryos, in contrast to recent results with stem cell cultures. In humans, degenerative changes have occurred as they approach the 14-day limit with the current methodology. In mice, embryos have been grown in vitro up to day 10.5 of development, the equivalent of about 4 weeks in humans, but this is obviously unethical in our species. So, the question is whether it is feasible to extend in vitro development of high-quality human embryos to this stage. However, even if it were, the extremely limited supply of such embryos means that any line of research requiring large numbers of cells would be extremely difficult to conduct and replicate in multiple laboratories (<a class="elsevierStyleCrossRef" href="#bb0215">Pera, 2017</a>).<a class="elsevierStyleCrossRef" href="#bb0105">Green et al. (2021)</a>, despite supporting human embryo research, are not very receptive to the recommendations of the International Society for Stem Cell Research (ISSCR) to extend the 14-day limit, as they consider that there are compelling reasons based on important biological events to maintain it, and believe that this institution should offer more convincing arguments for abandoning the current limit.Another group that is opposed to this extension is <a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger (2021)</a>. In response to McCully, they argue that the Warnock committee's reasons for the 14-day limit still apply despite advances in culturing human embryos in vitro. These authors believe that McCully seems unaware that such an extension is precisely the kind of slippery slope that opponents of the original law warned about. She also fails to take into account the argument based on the moral status of embryos: that only after the primitive line appears is the embryo a distinct individual and potential person. Moreover, research on embryos between 7 and 14 days old has only just begun to be exploited and it seems rash to extend it to 28 days. It is also not known how precisely post-implantation embryo culture would replicate the in utero environment, which may limit its applicability beyond 14 days.Therefore, several practical obstacles must first be overcome to facilitate this extended study. For example, conditions must be devised and improved to keep an embryo sustained and alive in a suitable environment (<a class="elsevierStyleCrossRef" href="#bb0005">Aach et al., 2017</a>). Although this challenge is difficult, it could be overcome with the help of 3D bioprinting and organoids (<a class="elsevierStyleCrossRef" href="#bb0005">Aach et al., 2017</a>; <a class="elsevierStyleCrossRef" href="#bb0040">Bredenoord et al., 2017</a>). To culture embryos at later developmental stages, such as beyond day 10.5 in mice or day 30 in macaques, it is essential to establish a functional placenta and a functional vascularised umbilical cord. This has been achieved artificially to help extremely premature lambs develop to near maturity (<a class="elsevierStyleCrossRef" href="#bb0205">Partridge et al., 2017</a>). Although this has not yet been achieved in humans, in the future, culture systems incorporated with a 3D printed vascularised umbilical cord and a functional artificial placenta in vitro could help extend in vitro embryo culture to later stages of development (<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al., 2021</a>).Ultimately, any decision to revise the 14-day rule must depend on whether the proposed changes can maintain the two main objectives of the rule: to support research and to accommodate various moral concerns (<a class="elsevierStyleCrossRef" href="#bb0150">Hyun et al., 2016</a>). It requires a careful assessment of the scientific feasibility and effective benefits of embryo research; it needs extensive research on public opinion regarding embryos, and a deliberative process that takes these elements into account. It does not need positions that only consider the beneficence of the research and its technical feasibility. This would be undemocratic and potentially a measure unsupported by a rigorous assessment of the science behind embryo research (<a class="elsevierStyleCrossRef" href="#bb0050">Cavaliere, 2017</a>).ConclusionsThe extended culture of embryos up to day 14 of development is leading to a better understanding of the events that take place between implantation and gastrulation. Despite this, it is necessary to improve the current culture conditions in order to optimise the technique and reach the maximum number of days allowed by law, as in most studies of this type, the embryos are blocked before day 14. Furthermore, it should be borne in mind that access to and availability of human embryos is limited, so other alternatives such as embryo development from stem cells should also be further explored.Once the culture conditions up to day 14 have been optimised, the idea of prolonging extended culture beyond day 14 is under debate, since with the current knowledge and with the help of existing technology, events such as gastrulation, hitherto unknown in humans, could be studied, allowing a better understanding of human embryonic development at later stages. In addition, this could be applied to areas such as assisted reproduction when making decisions on the choice of the best embryos or on the knowledge of diseases that may affect pregnancy and lead to miscarriage, for example. All of this with clear moral and ethical boundaries.FundingThis manuscript has not been funded by any organization.Conflict of interestThe authors declare that there is no conflict of interest regarding the publication of this manuscript." "textoCompletoSecciones" => array:1 [ "secciones" => array:12 [ 0 => array:3 [ "identificador" => "xres1786523" "titulo" => "Abstract" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "as0005" "titulo" => "Introduction" ] 1 => array:2 [ "identificador" => "as0010" "titulo" => "Materials and methods" ] 2 => array:2 [ "identificador" => "as0015" "titulo" => "Results and discussion" ] 3 => array:2 [ "identificador" => "as0020" "titulo" => "Conclusions" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec1566139" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "xpalclavsec1566141" "titulo" => "Abbreviations" ] 3 => array:3 [ "identificador" => "xres1786524" "titulo" => "Resumen" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "as0025" "titulo" => "Introducción" ] 1 => array:2 [ "identificador" => "as0030" "titulo" => "Materiales y métodos" ] 2 => array:2 [ "identificador" => "as0035" "titulo" => "Resultados y discusión" ] 3 => array:2 [ "identificador" => "as0040" "titulo" => "Conclusiones" ] ] ] 4 => array:2 [ "identificador" => "xpalclavsec1566140" "titulo" => "Palabras clave" ] 5 => array:2 [ "identificador" => "s0005" "titulo" => "Introduction" ] 6 => array:2 [ "identificador" => "s0010" "titulo" => "Materials and methods" ] 7 => array:3 [ "identificador" => "s0015" "titulo" => "Results and discussion" "secciones" => array:2 [ 0 => array:2 [ "identificador" => "s0020" "titulo" => "What is extended embryo culture up to 14 days and what are its conditions?" ] 1 => array:3 [ "identificador" => "s0025" "titulo" => "The conditions and components of the extended culture medium are explained below" "secciones" => array:8 [ 0 => array:2 [ "identificador" => "s0030" "titulo" => "Temperature" ] 1 => array:2 [ "identificador" => "s0035" "titulo" => "Gas environment" ] 2 => array:2 [ "identificador" => "s0040" "titulo" => "Culture media composition" ] 3 => array:2 [ "identificador" => "s0045" "titulo" => "What structures can we see with the culture up to day 14?" ] 4 => array:2 [ "identificador" => "s0050" "titulo" => "Stem cells and culture" ] 5 => array:2 [ "identificador" => "s0055" "titulo" => "Potential applications in embryology" ] 6 => array:2 [ "identificador" => "s0060" "titulo" => "14-day limit" ] 7 => array:2 [ "identificador" => "s0065" "titulo" => "So, should the 14-day limit be extended?" ] ] ] ] ] 8 => array:2 [ "identificador" => "s0070" "titulo" => "Conclusions" ] 9 => array:2 [ "identificador" => "s0075" "titulo" => "Funding" ] 10 => array:2 [ "identificador" => "s0080" "titulo" => "Conflict of interest" ] 11 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2022-02-23" "fechaAceptado" => "2022-05-19" "PalabrasClave" => array:2 [ "en" => array:2 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec1566139" "palabras" => array:5 [ 0 => "human embryo development" 1 => "in vitro embryo development" 2 => "pre-implantation" 3 => "in vitro embryo culture" 4 => "14-day-rule" ] ] 1 => array:4 [ "clase" => "abr" "titulo" => "Abbreviations" "identificador" => "xpalclavsec1566141" "palabras" => array:28 [ 0 => "2D" 1 => "3D" 2 => "ART" 3 => "ATP" 4 => "BSA" 5 => "CO2" 6 => "DMEM/F12" 7 => "DNA" 8 => "ESC" 9 => "GATA6" 10 => "HFE" 11 => "HSA" 12 => "ICM" 13 => "ISSCR" 14 => "ITS-X" 15 => "IVC1" 16 => "IVC2" 17 => "IVF" 18 => "Nanog" 19 => "NCBI" 20 => "NGS" 21 => "O2" 22 => "OCT4" 23 => "PKC" 24 => "SBF" 25 => "Sox17" 26 => "TSC" 27 => "XENC" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec1566140" "palabras" => array:5 [ 0 => "desarrollo embrionario humano" 1 => "desarrollo embrionario in vitro" 2 => "pre-implantación" 3 => "cultivo embrionario in vitro" 4 => "regla de los 14 días" ] ] ] ] "tieneResumen" => true "resumen" => array:2 [ "en" => array:3 [ "titulo" => "Abstract" "resumen" => "IntroductionThe early stages of human embryonic development prior to implantation have been extensively studied to improve reproductive outcomes. However, subsequent events are unknown. In vitro advances have allowed extended culture of embryos up to day 14, considered the legal limit. We aim to review aspects related culture conditions, morphogenesis events, potential applications of extended culture of human embryos as well as some ethical issues surrounding this topic. Materials and methodsThis literature review was carried out using different databases and websites by screening articles. Results and discussionExtended culture consists of culturing embryos up to day 14 of development. This requires specific culture conditions of temperature and gas concentration, as well as a medium composition that is different from that used in clinical practice in IVF laboratories. In addition to having made it possible to document in vitro the morphological events and structures of the embryo after implantation, the study of mosaic embryos at these hitherto hidden stages could provide valuable information for clinical practice. All this has been achieved within the 14-day limit, but due to scientific progress this can be considered restrictive and different reasons have been put forward why it should be extended. ConclusionsAlthough prolonged culture has provided a better understanding of the events occurring between implantation and gastrulation, culture conditions must be improved to optimise the technique, and once this is achieved, consideration should be given to extending this limit. Furthermore, alternatives to the use of embryos, such as the development of embryos from stem cells, should be further explored. All of this could have a future clinical application in terms of choosing the best embryos or finding out about diseases that affect pregnancy and lead to miscarriage." "secciones" => array:4 [ 0 => array:2 [ "identificador" => "as0005" "titulo" => "Introduction" ] 1 => array:2 [ "identificador" => "as0010" "titulo" => "Materials and methods" ] 2 => array:2 [ "identificador" => "as0015" "titulo" => "Results and discussion" ] 3 => array:2 [ "identificador" => "as0020" "titulo" => "Conclusions" ] ] ] "es" => array:3 [ "titulo" => "Resumen" "resumen" => "IntroducciónLas primeras etapas del desarrollo embrionario humano antes de la implantación han sido ampliamente estudiadas para mejorar los resultados reproductivos. Sin embargo, se desconocen los eventos posteriores. Los avances in vitro han permitido cultivar embriones hasta día 14, considerado el límite legal. Nuestro objetivo es hacer una revisión sobre las condiciones de cultivo, los eventos de morfogénesis, las posibles aplicaciones del cultivo extendido de embriones humanos, así como algunas cuestiones éticas que rodean este tema. Materiales y métodosEsta revisión bibliográfica se ha realizado empleando diferentes bases de datos y páginas web mediante el cribado de artículos. Resultados y discusiónEl cultivo extendido consiste en cultivar los embriones hasta día 14. Para ello se requieren unas condiciones de cultivo determinadas de temperatura y concentración de gases, así como una composición del medio que es diferente a la empleada en la práctica clínica de los laboratorios de FIV. Además de haber permitido documentar in vitro los eventos y estructuras morfológicas del embrión tras su implantación, el estudio de embriones mosaicos en estas etapas hasta ahora ocultas podría dar información valiosa para la práctica clínica. Todo esto se ha logrado dentro del límite de 14 días, pero debido al avance científico puede considerarse restrictivo, y se han expuesto diferentes motivos por los que debería ampliarse. ConclusionesAunque el cultivo prolongado ha permitido comprender mejor los acontecimientos que ocurren entre la implantación y la gastrulación, se deben mejorar las condiciones de cultivo para optimizar la técnica, y una vez conseguido, plantear la ampliación de este límite. Además, se deben seguir estudiando alternativas al uso de embriones como el desarrollo de embriones con células madre. Todo esto podría tener una aplicación clínica futura a la hora de elegir los mejores embriones o conocer enfermedades que afecten al embarazo y acaben en aborto." "secciones" => array:4 [ 0 => array:2 [ "identificador" => "as0025" "titulo" => "Introducción" ] 1 => array:2 [ "identificador" => "as0030" "titulo" => "Materiales y métodos" ] 2 => array:2 [ "identificador" => "as0035" "titulo" => "Resultados y discusión" ] 3 => array:2 [ "identificador" => "as0040" "titulo" => "Conclusiones" ] ] ] ] "multimedia" => array:4 [ 0 => array:8 [ "identificador" => "f0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1154 "Ancho" => 1778 "Tamanyo" => 153277 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "al0005" "detalle" => "Figure " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "Extended culture procedure up to day 14. The embryos are first thawed and then placed in culture medium for two hours for reexpansion. The next step is to remove the zona pellucida with tyrodes acid and then transfer them to the culture plate for culture until day 14 of development. Each day they can be observed under the microscope and their evolution can be evaluated." ] ] 1 => array:8 [ "identificador" => "f0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 674 "Ancho" => 1778 "Tamanyo" => 119460 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "al0010" "detalle" => "Figure " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "Events of human implantation morphogenesis. The embryo formed by the ICM (inner cell mass) and the trophectoderm is shown first. On the one hand, from the ICM, the epiblast and hypoblast segregate, forming the bilaminar disc. In the epiblast, a cavity called the pro-amniotic cavity begins to appear, which will later give rise to the amniotic cavity, and the hypoblast extends to form the yolk sac. On the other hand, the trophectoderm differentiates into cytotrophoblast and syncytiotrophoblast." ] ] 2 => array:8 [ "identificador" => "f0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 1012 "Ancho" => 1611 "Tamanyo" => 117760 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "al0015" "detalle" => "Figure " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "Non-integrated and integrated Stem cell-based embryo models. The upper figure of the schematic corresponds to an amniotic sac-like structure obtained from ESC. The second represents the elongated gastruloid with an anterior-posterior axis, also obtained from ESC. The third structure represents the embryoid, which has been obtained from ESC, XENC and TSC. And the last structure is known as the blastoid, made up of cells from ESC and TSC. (ESC: embryonic stem cells; XENC: extraembryonic endoderm cells; TSC: trophectoderm stem cells) (adapted from <a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam, 2021</a>)." ] ] 3 => array:8 [ "identificador" => "t0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "al0020" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:1 [ "tablatextoimagen" => array:1 [ 0 => array:1 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Advanced DMEM/F12 \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Concentration (mg/L) \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Concentration (mg/L) \t\t\t\t\t\t\n \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Non-essential amino acids: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Essential amino acids: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Glycine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">18.75 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Arginine hydrochloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">147.5 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Alanine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">4.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Histidine hydrochloride-H2O \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">31.48 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Asparagine-H2O \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Isoleucine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">54.47 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Aspartic acid \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">6.65 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Leucine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">59.05 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Cysteine hydrochloride-H2O \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.56 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Lysine hydrochloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">91.25 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Cysteine 2HCl \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">31.29 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Methionine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.24 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Glutamic acid \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.35 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Phenylalanine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">35.48 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Proline \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Threonine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">53.45 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Serine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">26.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Tryptophan \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">9.02 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Tyrosine disodium salt dihydrate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">55.79 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">L-Valine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">52.85 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Inorganic salts: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Vitamins: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Calcium chloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">116.6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Ascorbic acid phosphate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.5 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Cupric sulphate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.0013 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Biotin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.0035 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Ferric nitrate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Choline chloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">8.98 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Ferric sulphate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.417 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">D-Calcium pantothenate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.24 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Magnesium chloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">28.64 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Folic acid \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.65 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Magnesium sulphate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">48.84 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Niacinamide \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.02 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Potassium chloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">311.8 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Pyridoxine hydrochloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.0 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Sodium bicarbonate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2438.0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Riboflavin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.219 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Sodium chloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">6995.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Thiamine hydrochloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.17 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Sodium phosphate dibasic \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">71.02 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Vitamin B12 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.68 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Sodium phosphate monobasic \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">62.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">i-Inositol \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">132.6 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Zinc sulphate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.864 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Other components: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Ammonium metavanadate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.0E-4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">D-Glucose (Dextrose) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3151.0 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Manganous chloride \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5.0E-5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Ethanolamine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1.9 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Sodium selenite \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.005 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Hypoxanthine Na \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.39 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Proteins: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Linoleic acid \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.042 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">AlbuMAX® II \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">400.0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Lipoic acid \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.105 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Human Transferrin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Phenol red \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">8.1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Insulin Recombinant Full Chain \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10.0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Putrescine 2HCl \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.081 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Reducing agents: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Sodium pyruvate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">110.0 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Glutathione, monosodium \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1.0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Thymidine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.365 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">ITS-X \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Proteins: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Insulin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1000.0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Transferrin \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">550.0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Trace elements: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Sodium selenite \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.67 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Other components: \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Ethanolamine \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t">200.0 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] ] ] ] "descripcion" => array:1 [ "en" => "Composition and concentrations of basal medium Advanced DMEM/F12 and ITS-X. This table shows the different components that make up the basal medium Advanced DMEM/F12, as well as ITS-X, all with their respective concentrations in mg/L. Data obtained from <a class="elsevierStyleCrossRefs" href="#bb0285">ThermoFisher Scientific (2014a, 2014b)</a>." ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bs0005" "bibliografiaReferencia" => array:74 [ 0 => array:3 [ "identificador" => "bb0005" "etiqueta" => "Aach et al., 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Addressing the ethical issues raised by synthetic human entities with embryo-like features" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "J. Aach" 1 => "J. Lunshof" 2 => "E. Iyer" 3 => "G.M. Church" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:3 [ "tituloSerie" => "eLife Sci." "fecha" => "2017" "volumen" => "6" ] ] ] ] ] ] 1 => array:3 [ "identificador" => "bb0010" "etiqueta" => "Appleby and Bredenoord, 2018" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Should the 14-day rule for embryo research become the 28-day rule?" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "J.B. Appleby" 1 => "A.L. Bredenoord" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:4 [ "tituloSerie" => "EMBO Mol. Med." "fecha" => "2018" "volumen" => "10" "numero" => "9" ] ] ] ] ] ] 2 => array:3 [ "identificador" => "bb0015" "etiqueta" => "Baak et al., 2019" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Temperature of embryo culture for assisted reproduction" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "N.A. Baak" 1 => "A.E.P. Cantineau" 2 => "C. Farquhar" 3 => "D.R. Brison" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:5 [ "tituloSerie" => "Cochrane Database Syst. Rev." "fecha" => "2019" "volumen" => "9" "paginaInicial" => "1" "paginaFinal" => "34" ] ] ] ] ] ] 3 => array:3 [ "identificador" => "bb0020" "etiqueta" => "Beccari et al., 2018" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Multi-axialself-organization properties of mouse embryonic stem cells into gastruloids" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:9 [ 0 => "L. Beccari" 1 => "N. Moris" 2 => "M. Girgin" 3 => "D.A. Turner" 4 => "P. Baillie-Johnson" 5 => "A.C. Cossy" 6 => "M.P. Lutolf" 7 => "D. Duboule" 8 => "A.M. Arias" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat." "fecha" => "2018" "volumen" => "562" "numero" => "7726" "paginaInicial" => "272" "paginaFinal" => "276" ] ] ] ] ] ] 4 => array:3 [ "identificador" => "bb0025" "etiqueta" => "Bedzhov et al., 2014" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "In vitro culture of mouse blastocysts beyond the implantation stages" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "I. Bedzhov" 1 => "C.Y. Leung" 2 => "M. Bialecka" 3 => "M. Zernicka-Goetz" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/nprot.2014.186" "Revista" => array:7 [ "tituloSerie" => "Nat. Protoc." "fecha" => "2014" "volumen" => "9" "numero" => "12" "paginaInicial" => "2732" "paginaFinal" => "2739" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/25356584" "web" => "Medline" ] ] ] ] ] ] ] ] 5 => array:3 [ "identificador" => "bb0030" "etiqueta" => "Blackshaw and Rodger, 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Why we should not extend the 14-day rule" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "B.P. Blackshaw" 1 => "D. Rodger" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1136/medethics-2021-107317" "Revista" => array:7 [ "tituloSerie" => "J. Med. Ethics" "fecha" => "2021" "volumen" => "47" "numero" => "10" "paginaInicial" => "712" "paginaFinal" => "714" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/34112713" "web" => "Medline" ] ] ] ] ] ] ] ] 6 => array:3 [ "identificador" => "bb0035" "etiqueta" => "Bolton et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Mouse model of chromosome mosaicism reveals lineage-specific depletion of aneuploid cells and normal developmental potential" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:8 [ 0 => "H. Bolton" 1 => "S.J. Graham" 2 => "N. Van der Aa" 3 => "P. Kumar" 4 => "K. Theunis" 5 => "E. Fernandez Gallardo" 6 => "T. Voet" 7 => "M. Zernicka-Goetz" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat. Commun." "fecha" => "2016" "volumen" => "7" "numero" => "1" "paginaInicial" => "1" "paginaFinal" => "12" ] ] ] ] ] ] 7 => array:3 [ "identificador" => "bb0040" "etiqueta" => "Bredenoord et al., 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Human tissues in a dish: the research and ethical implications of organoid technology" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "A.L. Bredenoord" 1 => "H. Clevers" 2 => "J.A. Knoblich" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1126/science.aaf9414" "Revista" => array:6 [ "tituloSerie" => "Science." "fecha" => "2017" "volumen" => "355" "numero" => "6322" "paginaInicial" => "eaaf9414" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28104841" "web" => "Medline" ] ] ] ] ] ] ] ] 8 => array:3 [ "identificador" => "bb0045" "etiqueta" => "Castelyn, 2020" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Embryo experimentation: is there a case for moving beyond the ‘14-day rule’" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "G. Castelyn" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Monash Bioeth. Rev." "fecha" => "2020" "volumen" => "38" "numero" => "2" "paginaInicial" => "181" "paginaFinal" => "196" ] ] ] ] ] ] 9 => array:3 [ "identificador" => "bb0050" "etiqueta" => "Cavaliere, 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "A 14-day limit for bioethics: the debate over human embryo research" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "G. Cavaliere" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1186/s12910-016-0160-y" "Revista" => array:7 [ "tituloSerie" => "BMC Med. Ethics." "fecha" => "2017" "volumen" => "18" "numero" => "1" "paginaInicial" => "1" "paginaFinal" => "12" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28077127" "web" => "Medline" ] ] ] ] ] ] ] ] 10 => array:3 [ "identificador" => "bb0055" "etiqueta" => "Chattopadhyay et al., 2003" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Polyamines protect Escherichia coli cells from the toxic effect of oxygen" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "M.K. Chattopadhyay" 1 => "C.W. Tabor" 2 => "H. Tabor" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1073/pnas.2627990100" "Revista" => array:7 [ "tituloSerie" => "Proc. Natl. Acad. Sci. U. S. A." "fecha" => "2003" "volumen" => "100" "numero" => "5" "paginaInicial" => "2261" "paginaFinal" => "2265" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/12591940" "web" => "Medline" ] ] ] ] ] ] ] ] 11 => array:3 [ "identificador" => "bb0060" "etiqueta" => "Christianson et al., 2014" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Embryo catheter loading and embryo culture techniques: results of a worldwide web-based survey" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:8 [ 0 => "M.S. Christianson" 1 => "Y. Zhao" 2 => "G. Shoham" 3 => "I. Granot" 4 => "A. Safran" 5 => "A. Khafagy" 6 => "M. Leong" 7 => "Z. Shoham" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1007/s10815-014-0250-z" "Revista" => array:7 [ "tituloSerie" => "J. Assist. Reprod. Genet." "fecha" => "2014" "volumen" => "31" "numero" => "8" "paginaInicial" => "1029" "paginaFinal" => "1036" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/24913025" "web" => "Medline" ] ] ] ] ] ] ] ] 12 => array:3 [ "identificador" => "bb0065" "etiqueta" => "De Los Santos et al., 2013" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Reduced oxygen tension improves embryo quality but not clinical pregnancy rates: a randomized clinical study into ovum donation cycles" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:8 [ 0 => "M.J. De Los Santos" 1 => "P. Gámiz" 2 => "C. Albert" 3 => "A. Galán" 4 => "T. Viloria" 5 => "S. Pérez" 6 => "J.L. Romero" 7 => "J. Remohí" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.fertnstert.2013.03.044" "Revista" => array:7 [ "tituloSerie" => "Fertil. Steril." "fecha" => "2013" "volumen" => "100" "numero" => "2" "paginaInicial" => "402" "paginaFinal" => "407" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23628109" "web" => "Medline" ] ] ] ] ] ] ] ] 13 => array:3 [ "identificador" => "bb0070" "etiqueta" => "Deglincerti et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Self-organization of the in vitro attached human embryo" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:6 [ 0 => "A. Deglincerti" 1 => "G.F. Croft" 2 => "L.N. Pietila" 3 => "M. Zernicka-Goetz" 4 => "E.D. Siggia" 5 => "A.H. Brivanlou" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat." "fecha" => "2016" "volumen" => "533" "numero" => "7602" "paginaInicial" => "251" "paginaFinal" => "254" ] ] ] ] ] ] 14 => array:3 [ "identificador" => "bb0075" "etiqueta" => "Edwards et al., 1969" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Early stages of fertilization in vitro of human oocytes matured in vitro" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "R.G. Edwards" 1 => "B.D. Bavister" 2 => "P.C. Steptoe" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat." "fecha" => "1969" "volumen" => "221" "numero" => "5181" "paginaInicial" => "632" "paginaFinal" => "635" ] ] ] ] ] ] 15 => array:3 [ "identificador" => "bb0080" "etiqueta" => "Edwards et al., 1970" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Fertilization and cleavage in vitro of preovulator human oocytes" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "R.G. Edwards" 1 => "P.C. Steptoe" 2 => "J.M. Purdy" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat." "fecha" => "1970" "volumen" => "227" "numero" => "5265" "paginaInicial" => "1307" "paginaFinal" => "1309" ] ] ] ] ] ] 16 => array:3 [ "identificador" => "bb0085" "etiqueta" => "Enders et al., 1986" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Differentiation of the embryonic disc, amnion, and yolk sac in the rhesus monkey" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "A.C. Enders" 1 => "S. Schlafke" 2 => "A.G. Hendrickx" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1002/aja.1001770205" "Revista" => array:7 [ "tituloSerie" => "Am. J. Anat." "fecha" => "1986" "volumen" => "177" "numero" => "2" "paginaInicial" => "161" "paginaFinal" => "185" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/3788819" "web" => "Medline" ] ] ] ] ] ] ] ] 17 => array:3 [ "identificador" => "bb0090" "etiqueta" => "Etoc et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "A balance between secreted inhibitors and Edge sensing controls gastruloid self-organization" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:8 [ 0 => "F. Etoc" 1 => "J. Metzger" 2 => "A. Ruzo" 3 => "C. Kirst" 4 => "A. Yoney" 5 => "M.Z. Ozair" 6 => "A.H. Brivanlou" 7 => "E.D. Siggia" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.devcel.2016.09.016" "Revista" => array:7 [ "tituloSerie" => "Dev. Cell" "fecha" => "2016" "volumen" => "39" "numero" => "3" "paginaInicial" => "302" "paginaFinal" => "315" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27746044" "web" => "Medline" ] ] ] ] ] ] ] ] 18 => array:3 [ "identificador" => "bb0095" "etiqueta" => "Fürstova et al., 2008" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Comparison of the effect of individual saturated and unsaturated fatty acids on cell growth and death induction in the human pancreatic β-cell line NES2Y" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "V. Fürstova" 1 => "T. Kopska" 2 => "R.F. James" 3 => "J. Kovar" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.lfs.2007.12.023" "Revista" => array:7 [ "tituloSerie" => "Life Sci." "fecha" => "2008" "volumen" => "82" "numero" => "13-14" "paginaInicial" => "684" "paginaFinal" => "691" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/18272185" "web" => "Medline" ] ] ] ] ] ] ] ] 19 => array:3 [ "identificador" => "bb0100" "etiqueta" => "Greco et al., 2015" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Healthy babies after intrauterine transfer of mosaic aneuploid blastocysts" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "E. Greco" 1 => "M. Minasi" 2 => "F. Fiorentino" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1056/NEJMc1500421" "Revista" => array:7 [ "tituloSerie" => "N. Engl. J. Med." "fecha" => "2015" "volumen" => "373" "numero" => "21" "paginaInicial" => "2089" "paginaFinal" => "2090" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/26581010" "web" => "Medline" ] ] ] ] ] ] ] ] 20 => array:3 [ "identificador" => "bb0105" "etiqueta" => "Green et al., 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Don’t abandon 14-day limit on embryo research, it makes sense" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "R.M. Green" 1 => "M.D. West" 2 => "L. Hayflick" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/d41586-021-01595-7" "Revista" => array:6 [ "tituloSerie" => "Nature." "fecha" => "2021" "volumen" => "594" "numero" => "7863" "paginaInicial" => "333" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/34131341" "web" => "Medline" ] ] ] ] ] ] ] ] 21 => array:3 [ "identificador" => "bb0110" "etiqueta" => "Halloran et al., 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Arginine, agmatine, and polyamines: key regulators of conceptus development in mammals" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "K.M. Halloran" 1 => "C. Stenhouse" 2 => "G. Wu" 3 => "F.W. Bazer" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "LibroEditado" => array:6 [ "editores" => "G.Wu" "titulo" => "Amino Acids in Nutrition and Health" "paginaInicial" => "85" "paginaFinal" => "105" "serieTitulo" => "Advances in Experimental Medicine and Biology" "serieFecha" => "2021" ] ] ] ] ] ] 22 => array:3 [ "identificador" => "bb0115" "etiqueta" => "Harrison et al., 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Assembly of embryonic and extra-embryonic stem cells to mimic embryogeneis in vitro" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ 0 => "S.E. Harrison" 1 => "B. Sozen" 2 => "N. Christodoulou" 3 => "C. Kyprianou" 4 => "M. Zernicka-Goetz" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1126/science.aal1810" "Revista" => array:5 [ "tituloSerie" => "Sci." "fecha" => "2017" "volumen" => "356" "numero" => "6334" "paginaInicial" => "eaal1810" ] ] ] ] ] ] 23 => array:3 [ "identificador" => "bb0120" "etiqueta" => "Hertig et al., 1956" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "A description of 34 human ova within the first 17 days of development" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "A.T. Hertig" 1 => "J. Rock" 2 => "E.C. Adams" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1002/aja.1000980306" "Revista" => array:7 [ "tituloSerie" => "Am. J. Anat." "fecha" => "1956" "volumen" => "98" "numero" => "3" "paginaInicial" => "435" "paginaFinal" => "493" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/13362122" "web" => "Medline" ] ] ] ] ] ] ] ] 24 => array:3 [ "identificador" => "bb0125" "etiqueta" => "Hong et al., 2014" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Examining the temperature of embryo culture in in vitro fertilization: a randomized controlled trial comparing traditional core temperature (37°C) to a more physiologic, cooler temperature (36°C)" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ 0 => "K.H. Hong" 1 => "H. Lee" 2 => "E.J. Forman" 3 => "K.M. Upham" 4 => "R.T. Scott Jr." ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.fertnstert.2014.06.009" "Revista" => array:7 [ "tituloSerie" => "Fertil. Steril." "fecha" => "2014" "volumen" => "102" "numero" => "3" "paginaInicial" => "767" "paginaFinal" => "773" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/25044079" "web" => "Medline" ] ] ] ] ] ] ] ] 25 => array:3 [ "identificador" => "bb0130" "etiqueta" => "Human Fertilization and Embryology Act, 1990" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "UK Legislation, passed 1st November 1990. Her Majesty’s Stationery Office (HMSO)" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "Human Fertilization and Embryology Act" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Libro" => array:1 [ "fecha" => "1990" ] ] ] ] ] ] 26 => array:3 [ "identificador" => "bb0135" "etiqueta" => "Human Fertilization and Embryology Act, 2008" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "UK Legislation, passed 13th November 2008" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "Human Fertilization and Embryology Act" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "LibroEditado" => array:2 [ "titulo" => "Her Majesty’s Stationery Office (HMSO)" "serieFecha" => "2008" ] ] ] ] ] ] 27 => array:3 [ "identificador" => "bb0140" "etiqueta" => "Hurlbut et al., 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Revisiting the Warnock rule" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:15 [ 0 => "J.B. Hurlbut" 1 => "I. Hyun" 2 => "A.D. Levine" 3 => "R. Lovell-Badge" 4 => "J.E. Lunshof" 5 => "K.R. Matthews" 6 => "P. Mills" 7 => "A. Murdoch" 8 => "M.F. Pera" 9 => "C.T. Scott" 10 => "J. Tizzard" 11 => "M. Warnock" 12 => "M. Zernicka-Goetz" 13 => "Q. Zhou" 14 => "L. Zoloth" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/nbt.4015" "Revista" => array:7 [ "tituloSerie" => "Nat. Biotechnol." "fecha" => "2017" "volumen" => "35" "numero" => "11" "paginaInicial" => "1029" "paginaFinal" => "1042" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/29121021" "web" => "Medline" ] ] ] ] ] ] ] ] 28 => array:3 [ "identificador" => "bb0145" "etiqueta" => "Hussain et al., 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Exploring polyamines: functions in embryo/fetal development" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ 0 => "T. Hussain" 1 => "W. Ren" 2 => "N. Rahu" 3 => "D.H. Kalhoro" 4 => "Y. Yin" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.aninu.2016.12.002" "Revista" => array:7 [ "tituloSerie" => "Anim. Nutr." "fecha" => "2017" "volumen" => "3" "numero" => "1" "paginaInicial" => "7" "paginaFinal" => "10" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/29767087" "web" => "Medline" ] ] ] ] ] ] ] ] 29 => array:3 [ "identificador" => "bb0150" "etiqueta" => "Hyun et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Embryology policy: revisit the 14-day rule" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "I. Hyun" 1 => "A. Wilkerson" 2 => "J. Johnston" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat. N." "fecha" => "2016" "volumen" => "533" "numero" => "7602" "paginaInicial" => "169" "paginaFinal" => "171" ] ] ] ] ] ] 30 => array:3 [ "identificador" => "bb0155" "etiqueta" => "Hyun et al., 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Human embryo research beyond the primitive streak" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ 0 => "I. Hyun" 1 => "A.L. Bredenoord" 2 => "J. Briscoe" 3 => "S. Klipstein" 4 => "T. Tan" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1126/science.abf3751" "Revista" => array:7 [ "tituloSerie" => "Science." "fecha" => "2021" "volumen" => "371" "numero" => "6533" "paginaInicial" => "998" "paginaFinal" => "1000" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/33674483" "web" => "Medline" ] ] ] ] ] ] ] ] 31 => array:3 [ "identificador" => "bb0160" "etiqueta" => "Igarashi and Kashiwagi, 2000" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Polyamines: mysterious modulators of cellular functions" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "K. Igarashi" 1 => "K. Kashiwagi" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1006/bbrc.2000.2601" "Revista" => array:7 [ "tituloSerie" => "Biochem. Biophys. Res. Commun." "fecha" => "2000" "volumen" => "271" "numero" => "3" "paginaInicial" => "559" "paginaFinal" => "564" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/10814501" "web" => "Medline" ] ] ] ] ] ] ] ] 32 => array:3 [ "identificador" => "bb0165" "etiqueta" => "Kirkegaard et al., 2013" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "K. Kirkegaard" 1 => "J.J. Hindkjaer" 2 => "H.J. Ingerslev" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.fertnstert.2012.11.028" "Revista" => array:7 [ "tituloSerie" => "Fertil. Steril." "fecha" => "2013" "volumen" => "99" "numero" => "3" "paginaInicial" => "738" "paginaFinal" => "744" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23245683" "web" => "Medline" ] ] ] ] ] ] ] ] 33 => array:3 [ "identificador" => "bb0170" "etiqueta" => "Lusine, 2020" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "How good are we at modeling implantation?" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "A. Lusine" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1007/s10815-019-01667-8" "Revista" => array:7 [ "tituloSerie" => "J. Assist. Reprod. Genet." "fecha" => "2020" "volumen" => "37" "numero" => "1" "paginaInicial" => "17" "paginaFinal" => "19" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/31863287" "web" => "Medline" ] ] ] ] ] ] ] ] 34 => array:3 [ "identificador" => "bb0175" "etiqueta" => "McCully, 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "The time has come to extend the 14-day limit" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "S. McCully" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:5 [ "tituloSerie" => "J. Med. Ethics" "fecha" => "2021" "volumen" => "47" "numero" => "12" "paginaInicial" => "e66" ] ] ] ] ] ] 35 => array:3 [ "identificador" => "bb0180" "etiqueta" => "McLaren, 1984" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Where to draw the line" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "A. McLaren" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:5 [ "tituloSerie" => "P. Roy. Inst." "fecha" => "1984" "volumen" => "56" "paginaInicial" => "101" "paginaFinal" => "121" ] ] ] ] ] ] 36 => array:3 [ "identificador" => "bb0185" "etiqueta" => "Ménézo et al., 2013" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "New insights into human pre-implantation metabolism in vivo and in vitro" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "Y. Ménézo" 1 => "I. Lichtblau" 2 => "K. Elder" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1007/s10815-013-9953-9" "Revista" => array:7 [ "tituloSerie" => "J. Assist. Reprod. Genet." "fecha" => "2013" "volumen" => "30" "numero" => "3" "paginaInicial" => "293" "paginaFinal" => "303" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23430228" "web" => "Medline" ] ] ] ] ] ] ] ] 37 => array:3 [ "identificador" => "bb0190" "etiqueta" => "Nastri et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Low versus atmospheric oxygen tension for embryo culture in assisted reproduction: a systematic review and meta-analysis" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:9 [ 0 => "C.O. Nastri" 1 => "B.N. Nóbrega" 2 => "D.M. Teixeira" 3 => "J. Amorim" 4 => "L.M.M. Diniz" 5 => "M.W.P. Barbosa" 6 => "V.S.I. Giorgi" 7 => "V.N. Pileggi" 8 => "W.P. Martins" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.fertnstert.2016.02.037" "Revista" => array:7 [ "tituloSerie" => "Fertil. Steril." "fecha" => "2016" "volumen" => "106" "numero" => "1" "paginaInicial" => "95" "paginaFinal" => "104 e117" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27012651" "web" => "Medline" ] ] ] ] ] ] ] ] 38 => array:3 [ "identificador" => "bb0195" "etiqueta" => "Niu et al., 2019" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Dissecting primate early post-implantation development using long-term in vitro embryo culture" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:23 [ 0 => "Y. Niu" 1 => "N. Sun" 2 => "Y. Li" 3 => "Z. Huang" 4 => "J. Wu" 5 => "C. Si" 6 => "X. Dai" 7 => "C. Liu" 8 => "J. Wei" 9 => "L. Liu" 10 => "S. Feng" 11 => "Y. Kang" 12 => "W. Si" 13 => "H. Wang" 14 => "E. Zhang" 15 => "L. Zhao" 16 => "Z. Li" 17 => "X. Luo" 18 => "G. Cui" 19 => "G. Peng" 20 => "J.C. Izpisúa Belmonte" 21 => "W. Ji" 22 => "T. Tan" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1126/science.aaw5754" "Revista" => array:4 [ "tituloSerie" => "Sci." "fecha" => "2019" "volumen" => "366" "numero" => "6467" ] ] ] ] ] ] 39 => array:3 [ "identificador" => "bb0200" "etiqueta" => "Nuffield Council on Bioethics, 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Human embryo culture" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "Nuffield Council on Bioethics" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Libro" => array:3 [ "fecha" => "2017" "editorial" => "Nuffield Council on Bioethics" "editorialLocalizacion" => "London, UK" ] ] ] ] ] ] 40 => array:3 [ "identificador" => "bb0205" "etiqueta" => "Partridge et al., 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "An extra-uterine system to physiologically support the extreme premature lamb" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:18 [ 0 => "E.A. Partridge" 1 => "M.G. Davey" 2 => "M.A. Hornick" 3 => "P.E. McGovern" 4 => "A.Y. Mejaddam" 5 => "J.D. Vrecenak" 6 => "C. Mesas-Burgos" 7 => "A. Olive" 8 => "R.C. Caskey" 9 => "T.R. Weiland" 10 => "J. Han" 11 => "A.J. Schupper" 12 => "J.T. Connelly" 13 => "K.C. Dysart" 14 => "J. Rychik" 15 => "H.L. Hedrick" 16 => "W.H. Peranteau" 17 => "A.W. Flake" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/s41467-016-0009-6" "Revista" => array:7 [ "tituloSerie" => "Nat. Commun." "fecha" => "2017" "volumen" => "8" "numero" => "1" "paginaInicial" => "1" "paginaFinal" => "16" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28232747" "web" => "Medline" ] ] ] ] ] ] ] ] 41 => array:3 [ "identificador" => "bb0210" "etiqueta" => "Pawar and Jump, 2003" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Unsaturated fatty acid regulation of peroxisome proliferator-activated receptor α activity in rat primary hepatocytes" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "A. Pawar" 1 => "D.B. Jump" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1074/jbc.M306238200" "Revista" => array:7 [ "tituloSerie" => "J. Biol. Chem." "fecha" => "2003" "volumen" => "278" "numero" => "38" "paginaInicial" => "35931" "paginaFinal" => "35939" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/12853447" "web" => "Medline" ] ] ] ] ] ] ] ] 42 => array:3 [ "identificador" => "bb0215" "etiqueta" => "Pera, 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Human embryo research and the 14-day rule" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "M.F. Pera" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1242/dev.151191" "Revista" => array:7 [ "tituloSerie" => "Development." "fecha" => "2017" "volumen" => "144" "numero" => "11" "paginaInicial" => "1923" "paginaFinal" => "1925" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28559237" "web" => "Medline" ] ] ] ] ] ] ] ] 43 => array:3 [ "identificador" => "bb0220" "etiqueta" => "Popovic et al., 2019" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Extended in vitro culture of human embryos demonstrates the complex nature of diagnosing chromosomal mosaicism from a single trophectoderm biopsy" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:10 [ 0 => "M. Popovic" 1 => "L. Dhaenens" 2 => "J. Taelman" 3 => "A. Dheedene" 4 => "M. Bialecka" 5 => "P. De Sutter" 6 => "Chuva de Sousa" 7 => "S.M. Lopes" 8 => "B. Menten" 9 => "B. Heindryckx" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1093/humrep/dez012" "Revista" => array:7 [ "tituloSerie" => "Hum. Reprod." "fecha" => "2019" "volumen" => "34" "numero" => "4" "paginaInicial" => "758" "paginaFinal" => "769" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/30838420" "web" => "Medline" ] ] ] ] ] ] ] ] 44 => array:3 [ "identificador" => "bb0225" "etiqueta" => "Reimundo et al., 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Transferencia embrionaria única: estrategia clave para reducir el riesgo de embarazo múltiple en reproducción humana asistida" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "P. Reimundo" 1 => "J. Gutiérrez Romero" 2 => "T. Rodríguez Pérez" 3 => "E. Veiga" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Adv. Lab. Med." "fecha" => "2021" "volumen" => "2" "numero" => "2" "paginaInicial" => "189" "paginaFinal" => "198" ] ] ] ] ] ] 45 => array:3 [ "identificador" => "bb0230" "etiqueta" => "Rendón Abad, 2018" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Cultivo de preembriones humanos en presencia de alta tensión de oxígeno: resultados obstétricos y perinatales" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "M. Rendón Abad" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Libro" => array:2 [ "fecha" => "2018" "editorial" => "University of Valencia" ] ] ] ] ] ] 46 => array:3 [ "identificador" => "bb0235" "etiqueta" => "Rivron et al., 2018" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Blastocyst-like structures generated solely from stem cells" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:10 [ 0 => "N.C. Rivron" 1 => "J. Frias-Aldeguer" 2 => "E.J. Vrij" 3 => "J.C. Boisset" 4 => "J. Korving" 5 => "J. Vivie" 6 => "R.K. Truckenmuller" 7 => "A. van Oudenaarden" 8 => "C.A. van Blitterswijk" 9 => "N. Geijsen" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat." "fecha" => "2018" "volumen" => "557" "numero" => "7703" "paginaInicial" => "106" "paginaFinal" => "111" ] ] ] ] ] ] 47 => array:3 [ "identificador" => "bb0240" "etiqueta" => "Rossant, 2015" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Mouse and human blastocyst-derived stem cells: vive les differences" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "J. Rossant" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1242/dev.115451" "Revista" => array:7 [ "tituloSerie" => "Development." "fecha" => "2015" "volumen" => "142" "numero" => "1" "paginaInicial" => "9" "paginaFinal" => "12" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/25516964" "web" => "Medline" ] ] ] ] ] ] ] ] 48 => array:3 [ "identificador" => "bb0245" "etiqueta" => "Rossant and Tam, 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Opportunities and challenges with stem cell-based embryo models" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "J. Rossant" 1 => "P.P. Tam" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Stem Cell Rep." "fecha" => "2021" "volumen" => "16" "numero" => "5" "paginaInicial" => "1031" "paginaFinal" => "1038" ] ] ] ] ] ] 49 => array:3 [ "identificador" => "bb0250" "etiqueta" => "Shahbazi and Zernicka-Goetz, 2018" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Deconstructing and reconstructing the mouse and human early embryo" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "M. Shahbazi" 1 => "M. Zernicka-Goetz" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/s41556-018-0144-x" "Revista" => array:7 [ "tituloSerie" => "Nat. Cell Biol." "fecha" => "2018" "volumen" => "20" "numero" => "8" "paginaInicial" => "878" "paginaFinal" => "887" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/30038253" "web" => "Medline" ] ] ] ] ] ] ] ] 50 => array:3 [ "identificador" => "bb0255" "etiqueta" => "Shahbazi et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Self-organization of the human embryo in the absence of maternal tissues" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:14 [ 0 => "M.N. Shahbazi" 1 => "A. Jedrusik" 2 => "S. Vuoristo" 3 => "G. Recher" 4 => "A. Hupalowska" 5 => "V. Bolton" 6 => "N.M.E. Fogarty" 7 => "A. Campbell" 8 => "L.G. Devito" 9 => "D. Ilic" 10 => "Y. Khalaf" 11 => "K.K. Niakan" 12 => "S. Fishel" 13 => "M. Zernicka-Goetz" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/ncb3347" "Revista" => array:7 [ "tituloSerie" => "Nat. Cell Biol." "fecha" => "2016" "volumen" => "18" "numero" => "6" "paginaInicial" => "700" "paginaFinal" => "708" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27144686" "web" => "Medline" ] ] ] ] ] ] ] ] 51 => array:3 [ "identificador" => "bb0260" "etiqueta" => "Shahbazi et al., 2020" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Developmental potential of aneuploid human embryos cultured beyond implantation" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:12 [ 0 => "M.N. Shahbazi" 1 => "T. Wang" 2 => "X. Tao" 3 => "B.A. Weatherbee" 4 => "L. Sun" 5 => "Y. Zhan" 6 => "L. Keller" 7 => "G.D. Smith" 8 => "A. Pellicer" 9 => "R.T. Scott" 10 => "E. Seli" 11 => "M. Zernicka-Goetz" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/s41467-019-13993-7" "Revista" => array:7 [ "tituloSerie" => "Nat. Commun." "fecha" => "2020" "volumen" => "11" "numero" => "1" "paginaInicial" => "1" "paginaFinal" => "15" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/31911652" "web" => "Medline" ] ] ] ] ] ] ] ] 52 => array:3 [ "identificador" => "bb0265" "etiqueta" => "Shao et al., 2017" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Self-organized amniogenesis by human pluripotent stem cells in a biomimetic implantation-like niche" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:10 [ 0 => "Y. Shao" 1 => "K. Taniguchi" 2 => "K. Gurdziel" 3 => "R.F. Townshend" 4 => "X. Xue" 5 => "K.M. Yong" 6 => "J. Sang" 7 => "J.R. Spence" 8 => "D.L. Gumucio" 9 => "J. Fu" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/nmat4829" "Revista" => array:7 [ "tituloSerie" => "Nat. Mater." "fecha" => "2017" "volumen" => "16" "numero" => "4" "paginaInicial" => "419" "paginaFinal" => "425" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27941807" "web" => "Medline" ] ] ] ] ] ] ] ] 53 => array:3 [ "identificador" => "bb0270" "etiqueta" => "Sozen et al., 2018" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Self-assembly of embryonic and two extra-embryonic stem cell types into gastrulating embryo-like structures" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:11 [ 0 => "B. Sozen" 1 => "G. Amadei" 2 => "A. Cox" 3 => "R. Wang" 4 => "Czukiewska Ellen" 5 => "L. Chappell" 6 => "T. Voet" 7 => "G. Michel" 8 => "N. Jing" 9 => "D.M. Glover" 10 => "M. Zernicka-Goetz" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/s41556-018-0147-7" "Revista" => array:7 [ "tituloSerie" => "Nat. Cell Biol." "fecha" => "2018" "volumen" => "20" "numero" => "8" "paginaInicial" => "979" "paginaFinal" => "989" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/30038254" "web" => "Medline" ] ] ] ] ] ] ] ] 54 => array:3 [ "identificador" => "bb0275" "etiqueta" => "Steptoe and Edwards, 1978" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Birth after the reimplantation of a human embryo" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "P.C. Steptoe" 1 => "R.G. Edwards" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:5 [ "tituloSerie" => "Lancet." "fecha" => "1978" "volumen" => "312" "numero" => "8085" "paginaInicial" => "366" ] ] ] ] ] ] 55 => array:3 [ "identificador" => "bb0280" "etiqueta" => "Sunde et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Time to take human embryo culture seriously" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:8 [ 0 => "A. Sunde" 1 => "D. Brison" 2 => "J. Dumoulin" 3 => "J. Harper" 4 => "K. Lundin" 5 => "M.C. Magli" 6 => "E. van den Abbeel" …1 ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1093/humrep/dew157" "Revista" => array:7 [ "tituloSerie" => "Hum. Reprod." "fecha" => "2016" "volumen" => "31" "numero" => "10" "paginaInicial" => "2174" "paginaFinal" => "2182" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 56 => array:3 [ "identificador" => "bb0285" "etiqueta" => "ThermoFisher Scientific, 2014a" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Advanced DMEM/F-12" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ …1] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "WWW" => array:2 [ "link" => "https://www.thermofisher.com/order/catalog/product/12634010" "fecha" => "2014" ] ] ] ] ] ] 57 => array:3 [ "identificador" => "bb0290" "etiqueta" => "ThermoFisher Scientific, 2014b" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Insulin-Transferrin-Selenium-Ethaolamine (ITS-X) (100X)" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ …1] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "WWW" => array:2 [ "link" => "https://www.thermofisher.com/order/catalog/product/51500056" "fecha" => "2014" ] ] ] ] ] ] 58 => array:3 [ "identificador" => "bb0295" "etiqueta" => "ThermoFisher Scientific, 2014c" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "AlbuMAX® I and AlbuMAX® II" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ …1] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "WWW" => array:2 [ "link" => "https://www.thermofisher.com/document-connect/document-connect.html?url=https%3A%2F%2Fassets.thermofisher.com%2FTFS-Assets%2FLSG%2Fmanuals%2F3117.pdf" "fecha" => "2014" ] ] ] ] ] ] 59 => array:3 [ "identificador" => "bb0300" "etiqueta" => "ThermoFisher Scientific, 2020a" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Knockout™ SR User Guide" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ …1] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "WWW" => array:2 [ "link" => "https://www.thermofisher.com/order/catalog/product/10828010?SID=srch-srp-10828010" "fecha" => "2020" ] ] ] ] ] ] 60 => array:3 [ "identificador" => "bb0305" "etiqueta" => "ThermoFisher Scientific, 2020b" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "GlutaMAX™ Product Information Sheet" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ …1] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "WWW" => array:2 [ "link" => "https://assets.fishersci.com/TFS-Assets/LSG/manuals/GlutaMAX_PI.pdf?_ga=2.254523471.695974732.1645097875-1058616482.1617892711&_gac=1.224585576.1642763480.CjwKCAiA0KmPBhBqEiwAJqKK49EYq0AMKeAS6erwSS_dE8rEgLEcD9A1li5aUGnLL9hUWNG-7ItWeBoCRMcQAvD_BwE" "fecha" => "2020" ] ] ] ] ] ] 61 => array:3 [ "identificador" => "bb0310" "etiqueta" => "van den Brink et al., 2014" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Symmetry breaking, germ layer specification and axial organisation in aggregates of mouse embryonic stem cells" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:7 [ …7] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1242/dev.113001" "Revista" => array:7 [ "tituloSerie" => "Development." "fecha" => "2014" "volumen" => "141" "numero" => "22" "paginaInicial" => "4231" "paginaFinal" => "4242" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 62 => array:3 [ "identificador" => "bb0315" "etiqueta" => "van der Valk et al., 2018" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Fetal Bovine Serum (FBS): Past-Present-Future" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:17 [ …17] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Altex." "fecha" => "2018" "volumen" => "35" "numero" => "1" "paginaInicial" => "1" "paginaFinal" => "20" ] ] ] ] ] ] 63 => array:3 [ "identificador" => "bb0320" "etiqueta" => "van Montfoort et al., 2020" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Reduced oxygen concentration during human IVF culture improves embryo utilization and cumulative pregnancy rates per cycle" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:7 [ …7] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1093/hropen/hoz036" "Revista" => array:6 [ "tituloSerie" => "Hum. Reprod. Open." "fecha" => "2020" "volumen" => "2020" "numero" => "1" "paginaInicial" => "hoz036" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 64 => array:3 [ "identificador" => "bb0325" "etiqueta" => "Wale and Gardner, 2010" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Time-lapse analysis of mouse embryo development in oxygen gradients" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ …2] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.rbmo.2010.04.028" "Revista" => array:7 [ "tituloSerie" => "Reprod. BioMed. Online" "fecha" => "2010" "volumen" => "21" "numero" => "3" "paginaInicial" => "402" "paginaFinal" => "410" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 65 => array:3 [ "identificador" => "bb0330" "etiqueta" => "Wang et al., 2002" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Vitamin C and vitamin E supplementation reduce oxidative stress–induced embryo toxicity and improve the blastocyst development rate" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:6 [ …6] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/s0015-0282(02)04236-x" "Revista" => array:7 [ "tituloSerie" => "Fertil. Steril." "fecha" => "2002" "volumen" => "78" "numero" => "6" "paginaInicial" => "1272" "paginaFinal" => "1277" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 66 => array:3 [ "identificador" => "bb0335" "etiqueta" => "Warnock, 1984" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Report of the Committee of Inquiry into Human Fertilization and Embryology" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ …1] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Libro" => array:3 [ "fecha" => "1984" "editorial" => "Her Majesty’s Stationery Office (HMSO)" "editorialLocalizacion" => "London" ] ] ] ] ] ] 67 => array:3 [ "identificador" => "bb0340" "etiqueta" => "Weimar et al., 2013" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "In-vitro model systems for the study of human embryo-endometrium interactions" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ …5] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.rbmo.2013.08.002" "Revista" => array:6 [ "tituloSerie" => "Reprod. BioMed. Online" "fecha" => "2013" "volumen" => "27" "numero" => "5" "paginaInicial" => "461.-476" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 68 => array:3 [ "identificador" => "bb0345" "etiqueta" => "Williams and Johnson, 2020" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Adapting the 14-day rule for embryo research to encompass evolving technologies" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ …2] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.rbms.2019.12.002" "Revista" => array:6 [ "tituloSerie" => "Reprod. Biomed. Soc. Online." "fecha" => "2020" "volumen" => "10" "paginaInicial" => "1" "paginaFinal" => "9" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 69 => array:3 [ "identificador" => "bb0350" "etiqueta" => "Wu and Morris Jr, 1998" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Arginine metabolism: nitric oxide and beyond" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ …2] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1042/bj3360001" "Revista" => array:7 [ "tituloSerie" => "Biochem. J." "fecha" => "1998" "volumen" => "336" "numero" => "1" "paginaInicial" => "1" "paginaFinal" => "17" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 70 => array:3 [ "identificador" => "bb0355" "etiqueta" => "Zander-Fox et al., 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Albumin used in human IVF contain different levels of lipids and modify embryo and fetal growth in a mouse model" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ …3] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1007/s10815-021-02255-5" "Revista" => array:6 [ "tituloSerie" => "J. Assist. Reprod. Genet." "fecha" => "2021" "volumen" => "38" "paginaInicial" => "2371" "paginaFinal" => "2381" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 71 => array:3 [ "identificador" => "bb0360" "etiqueta" => "Zernicka-Goetz et al., 2016" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Culture of Human Embryos through Implantation Stages in Vitro" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ …4] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Libro" => array:3 [ "fecha" => "2016" "paginaInicial" => "1" "paginaFinal" => "7" ] ] ] ] ] ] 72 => array:3 [ "identificador" => "bb0365" "etiqueta" => "Zhai et al., 2021" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Human embryonic development: from peri-implantation to gastrulation" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ …4] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.tcb.2021.07.008" "Revista" => array:7 [ "tituloSerie" => "Trends Cell Biol." "fecha" => "2021" "volumen" => "32" "numero" => "1" "paginaInicial" => "18" "paginaFinal" => "29" "link" => array:1 [ 0 => array:2 [ …2] ] ] ] ] ] ] ] 73 => array:3 [ "identificador" => "bb0370" "etiqueta" => "Zheng et al., 2019" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Controlled modelling of human epiblast and amnion development using stem cells" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:11 [ …11] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Nat." "fecha" => "2019" "volumen" => "573" "numero" => "7774" "paginaInicial" => "421" "paginaFinal" => "425" ] ] ] ] ] ] ] ] ] ] ] "idiomaDefecto" => "en" "url" => "/23409320/0000000900000003/v1_202210200635/S2340932022000068/v1_202210200635/en/main.assets" "Apartado" => array:4 [ "identificador" => "93980" "tipo" => "SECCION" "en" => array:2 [ "titulo" => "Revisiones" "idiomaDefecto" => true ] "idiomaDefecto" => "en" ] "PDF" => "https://static.elsevier.es/multimedia/23409320/0000000900000003/v1_202210200635/S2340932022000068/v1_202210200635/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S2340932022000068?idApp=UINPBA00004N"]

Statistics

Review

Extended embryo culture up to 14 days

Cultivo embrionario extendido hasta 14 días

David Ortega-Jaena, Angel Martinb, M.L. Pardiñasb, Amparo Mifsuda, Amparo Mercadera, M.J. de los Santosa,

Corresponding author

mariajose.delossantos@ivirma.com

Corresponding author at: Laboratorio de FIV, Instituto Valenciano de Infertilidad, IVI-RMA, Pl. de la Policía Local, 3, Valencia 46015, Spain.

a Laboratorio FIV, Instituto Valenciano de Infertilidad, IVI-RMA, Valencia, Spain

b Fundación IVI, Valencia, Spain

Read

643

Times
was read the article

100

Total PDF

543

Total HTML

Share statistics

 array:23 [
  "pii" => "S2340932022000068"
  "issn" => "23409320"
  "doi" => "10.1016/j.medre.2022.100118"
  "estado" => "S300"
  "fechaPublicacion" => "2022-09-01"
  "aid" => "100118"
  "copyright" => "Asociaci&#243;n para el Estudio de la Biolog&#237;a de la Reproducci&#243;n y Sociedad Espa&#241;ola de Fertilidad"
  "copyrightAnyo" => "2022"
  "documento" => "article"
  "crossmark" => 1
  "subdocumento" => "rev"
  "cita" => "Medicina Reproductiva y Embriolog&#237;a Cl&#237;nica. 2022;9:"
  "abierto" => array:3 [
    "ES" => false
    "ES2" => false
    "LATM" => false
  ]
  "gratuito" => false
  "lecturas" => array:1 [
    "total" => 0
  ]
  "itemSiguiente" => array:18 [
    "pii" => "S2340932022000093"
    "issn" => "23409320"
    "doi" => "10.1016/j.medre.2022.100121"
    "estado" => "S300"
    "fechaPublicacion" => "2022-09-01"
    "aid" => "100121"
    "copyright" => "Asociaci&#243;n para el Estudio de la Biolog&#237;a de la Reproducci&#243;n y Sociedad Espa&#241;ola de Fertilidad"
    "documento" => "article"
    "crossmark" => 1
    "subdocumento" => "rev"
    "cita" => "Medicina Reproductiva y Embriolog&#237;a Cl&#237;nica. 2022;9:"
    "abierto" => array:3 [
      "ES" => false
      "ES2" => false
      "LATM" => false
    ]
    "gratuito" => false
    "lecturas" => array:1 [
      "total" => 0
    ]
    "en" => array:11 [
      "idiomaDefecto" => true
      "cabecera" => "<span class="elsevierStyleTextfn">REVIEW</span>"
      "titulo" => "Sperm DNA fragmentation and microfluidics&#58; A new era in human sperm selection"
      "tienePdf" => "en"
      "tieneTextoCompleto" => "en"
      "tieneResumen" => array:2 [
        0 => "en"
        1 => "es"
      ]
      "contieneResumen" => array:2 [
        "en" => true
        "es" => true
      ]
      "contieneTextoCompleto" => array:1 [
        "en" => true
      ]
      "contienePdf" => array:1 [
        "en" => true
      ]
      "resumenGrafico" => array:2 [
        "original" => 0
        "multimedia" => array:8 [
          "identificador" => "f0005"
          "etiqueta" => "Fig&#46; 1"
          "tipo" => "MULTIMEDIAFIGURA"
          "mostrarFloat" => true
          "mostrarDisplay" => false
          "figura" => array:1 [
            0 => array:4 [
              "imagen" => "gr1.jpeg"
              "Alto" => 4159
              "Ancho" => 2953
              "Tamanyo" => 618547
            ]
          ]
          "detalles" => array:1 [
            0 => array:3 [
              "identificador" => "al0005"
              "detalle" => "Fig&#46; "
              "rol" => "short"
            ]
          ]
          "descripcion" => array:1 [
            "en" => "<p id="sp0005" class="elsevierStyleSimplePara elsevierViewall">Microfluidic-based devices for sperm selection&#46; A<span class="elsevierStyleHsp" style=""></span>&#124; First device based on sperm motility&#46; It is consisted of 2-inlet and 2-outlet linked by channels&#46; The exerted flow runs parallel to the passive sample current&#44; which allows sorting of motile sperm in a separate chamber&#46; B<span class="elsevierStyleHsp" style=""></span>&#124; Device based on sperm rheotaxis with 3 reservoirs and 4 channels&#46; A flow is generated by hydrostatic pressure allowing the selection of sperm that swims against it&#46; C<span class="elsevierStyleHsp" style=""></span>&#124; Microfluidic sorting device composed of two chambers connected to a micropump which generates a flow to select sperm according to rheotaxis behavior&#46; D<span class="elsevierStyleHsp" style=""></span>&#124; Device consisting of a pool surrounded by U-shape channels&#46; Different progesterone concentrations are added at inlet A while motile sperm is sorted at outlet B&#46; E<span class="elsevierStyleHsp" style=""></span>&#124; Represents the CS-10 designed with an electric field which allows the transport of the motile sperm across a membrane&#46; F<span class="elsevierStyleHsp" style=""></span>&#124; System that selects the motile sperm swimming fastest through a passive flow&#46; G<span class="elsevierStyleHsp" style=""></span>&#124; Passive mechanism composed of a porous membrane capable of filtering sperm with better motility and fragmentation&#46; H<span class="elsevierStyleHsp" style=""></span>&#124; The hydrodynamic behavior and swimming direction of the sperm is simulated in the SPARTAN for selection&#46; It is made of 2 chambers connected by a channel composed of spaced micropillars&#46; I<span class="elsevierStyleHsp" style=""></span>&#124; Based on boundary-following behavior&#44; the selection in this device occurred by tracking sperm in a circular space of 500 microchannels&#46; J<span class="elsevierStyleHsp" style=""></span>&#124; In this device&#44; a patterned layer is placed forming a semi-circular structure while a covering layer is superimposed to establish inlet and outlet chambers to collect the highest quality sperm&#46; K<span class="elsevierStyleHsp" style=""></span>&#124; The commercially available microchip is composed of 2 wells separated by a membrane with the capacity to filter sperm with better motility and fragmentation during a short incubation period&#46; White sperm&#58; non-motile&#46; Blue sperm&#58; motile&#46; Green arrows indicate the direction of flow and red ones magnify the selected section&#46; Adapted from A&#41; <a class="elsevierStyleCrossRef" href="#bb0135">Cho et al&#46;&#44; 2003</a> B&#41; <a class="elsevierStyleCrossRef" href="#bb0535">Seo et al&#46;&#44; 2007</a> C&#41; <a class="elsevierStyleCrossRef" href="#bb0700">Zhang et al&#46;&#44; 2015</a>a D&#41; <a class="elsevierStyleCrossRef" href="#bb0705">Zhang et al&#46;&#44; 2015b</a> E&#41; <a class="elsevierStyleCrossRef" href="#bb0030">Ainsworth et al&#46;&#44; 2005</a>&#8239;F&#41; <a class="elsevierStyleCrossRef" href="#bb0695">Zhang et al&#46;&#44; 2011</a> G&#41; <a class="elsevierStyleCrossRef" href="#bb0060">Asghar et al&#46;&#44; 2014</a> H&#41; <a class="elsevierStyleCrossRef" href="#bb0130">Chinnasamy et al&#46;&#44; 2018</a> I&#41; <a class="elsevierStyleCrossRef" href="#bb0400">Nosrati et al&#46;&#44; 2014</a>&#8239;J&#41; <a class="elsevierStyleCrossRef" href="#bb0675">Xiao et al&#46;&#44; 2021</a> K&#41; <a class="elsevierStyleCrossRef" href="#bb0375">Meseguer et al&#46;&#44; 2021</a>&#46; &#40;For interpretation of the references to color in this figure legend&#44; the reader is referred to the web version of this article&#46;&#41;</p>"
          ]
        ]
      ]
      "autores" => array:1 [
        0 => array:2 [
          "autoresLista" => "Maria Luisa Pardi&#241;as, Angel Martin, David Ortega-Ja&#233;n, Jose Mar&#237;a De los Santos, Thamara Viloria, Pilar Gamiz, Maria Jos&#233; De los Santos"
          "autores" => array:7 [
            0 => array:2 [
              "nombre" => "Maria Luisa"
              "apellidos" => "Pardi&#241;as"
            ]
            1 => array:2 [
              "nombre" => "Angel"
              "apellidos" => "Martin"
            ]
            2 => array:2 [
              "nombre" => "David"
              "apellidos" => "Ortega-Ja&#233;n"
            ]
            3 => array:2 [
              "nombre" => "Jose Mar&#237;a"
              "apellidos" => "De los Santos"
            ]
            4 => array:2 [
              "nombre" => "Thamara"
              "apellidos" => "Viloria"
            ]
            5 => array:2 [
              "nombre" => "Pilar"
              "apellidos" => "Gamiz"
            ]
            6 => array:2 [
              "nombre" => "Maria Jos&#233;"
              "apellidos" => "De los Santos"
            ]
          ]
        ]
      ]
    ]
    "idiomaDefecto" => "en"
    "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S2340932022000093?idApp=UINPBA00004N"
    "url" => "/23409320/0000000900000003/v1_202210200635/S2340932022000093/v1_202210200635/en/main.assets"
  ]
  "itemAnterior" => array:18 [
    "pii" => "S234093202200007X"
    "issn" => "23409320"
    "doi" => "10.1016/j.medre.2022.100119"
    "estado" => "S300"
    "fechaPublicacion" => "2022-09-01"
    "aid" => "100119"
    "copyright" => "Asociaci&#243;n para el Estudio de la Biolog&#237;a de la Reproducci&#243;n y Sociedad Espa&#241;ola de Fertilidad"
    "documento" => "article"
    "crossmark" => 1
    "subdocumento" => "rev"
    "cita" => "Medicina Reproductiva y Embriolog&#237;a Cl&#237;nica. 2022;9:"
    "abierto" => array:3 [
      "ES" => false
      "ES2" => false
      "LATM" => false
    ]
    "gratuito" => false
    "lecturas" => array:1 [
      "total" => 0
    ]
    "es" => array:11 [
      "idiomaDefecto" => true
      "cabecera" => "<span class="elsevierStyleTextfn">Revisi&#243;n</span>"
      "titulo" => "Aplicaci&#243;n de la inteligencia artificial en el laboratorio de reproducci&#243;n asistida&#46; Trabajo de revisi&#243;n"
      "tienePdf" => "es"
      "tieneTextoCompleto" => "es"
      "tieneResumen" => array:2 [
        0 => "es"
        1 => "en"
      ]
      "contieneResumen" => array:2 [
        "es" => true
        "en" => true
      ]
      "contieneTextoCompleto" => array:1 [
        "es" => true
      ]
      "contienePdf" => array:1 [
        "es" => true
      ]
      "resumenGrafico" => array:2 [
        "original" => 0
        "multimedia" => array:8 [
          "identificador" => "f0005"
          "etiqueta" => "Figura 1"
          "tipo" => "MULTIMEDIAFIGURA"
          "mostrarFloat" => true
          "mostrarDisplay" => false
          "figura" => array:1 [
            0 => array:4 [
              "imagen" => "gr1.jpeg"
              "Alto" => 1056
              "Ancho" => 1903
              "Tamanyo" => 115689
            ]
          ]
          "detalles" => array:1 [
            0 => array:3 [
              "identificador" => "al0005"
              "detalle" => "Figura "
              "rol" => "short"
            ]
          ]
          "descripcion" => array:1 [
            "es" => "<p id="sp0005" class="elsevierStyleSimplePara elsevierViewall">Esquema simplificado del proceso de laboratorio de reproducci&#243;n asistida&#46;</p> <p id="sp0010" class="elsevierStyleSimplePara elsevierViewall">El embri&#243;logo realiza un an&#225;lisis de semen del paciente masculino y eval&#250;a los ovocitos de la paciente&#46; En el caso de la t&#233;cnica ICSI&#44; se selecciona el mejor espermatozoide para inyectar en el ovocito&#46; Durante el desarrollo embrionario se selecciona el embri&#243;n que se va a transferir y los embriones que se van a conservar mediante vitrificaci&#243;n&#44; se descartan los embriones de desecho&#46;</p>"
          ]
        ]
      ]
      "autores" => array:1 [
        0 => array:2 [
          "autoresLista" => "Paula Mart&#237;n-Climent, Juan M&#46; Moreno-Garc&#237;a"
          "autores" => array:2 [
            0 => array:2 [
              "nombre" => "Paula"
              "apellidos" => "Mart&#237;n-Climent"
            ]
            1 => array:2 [
              "nombre" => "Juan M&#46;"
              "apellidos" => "Moreno-Garc&#237;a"
            ]
          ]
        ]
      ]
    ]
    "idiomaDefecto" => "es"
    "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S234093202200007X?idApp=UINPBA00004N"
    "url" => "/23409320/0000000900000003/v1_202210200635/S234093202200007X/v1_202210200635/es/main.assets"
  ]
  "en" => array:18 [
    "idiomaDefecto" => true
    "cabecera" => "<span class="elsevierStyleTextfn">Review</span>"
    "titulo" => "Extended embryo culture up to 14 days"
    "tieneTextoCompleto" => true
    "autores" => array:1 [
      0 => array:4 [
        "autoresLista" => "David Ortega-Jaen, Angel Martin, M&#46;L&#46; Pardi&#241;as, Amparo Mifsud, Amparo Mercader, M&#46;J&#46; de los Santos"
        "autores" => array:6 [
          0 => array:3 [
            "nombre" => "David"
            "apellidos" => "Ortega-Jaen"
            "referencia" => array:1 [
              0 => array:2 [
                "etiqueta" => "<span class="elsevierStyleSup">a</span>"
                "identificador" => "af0005"
              ]
            ]
          ]
          1 => array:3 [
            "nombre" => "Angel"
            "apellidos" => "Martin"
            "referencia" => array:1 [
              0 => array:2 [
                "etiqueta" => "<span class="elsevierStyleSup">b</span>"
                "identificador" => "af0010"
              ]
            ]
          ]
          2 => array:3 [
            "nombre" => "M&#46;L&#46;"
            "apellidos" => "Pardi&#241;as"
            "referencia" => array:1 [
              0 => array:2 [
                "etiqueta" => "<span class="elsevierStyleSup">b</span>"
                "identificador" => "af0010"
              ]
            ]
          ]
          3 => array:3 [
            "nombre" => "Amparo"
            "apellidos" => "Mifsud"
            "referencia" => array:1 [
              0 => array:2 [
                "etiqueta" => "<span class="elsevierStyleSup">a</span>"
                "identificador" => "af0005"
              ]
            ]
          ]
          4 => array:3 [
            "nombre" => "Amparo"
            "apellidos" => "Mercader"
            "referencia" => array:1 [
              0 => array:2 [
                "etiqueta" => "<span class="elsevierStyleSup">a</span>"
                "identificador" => "af0005"
              ]
            ]
          ]
          5 => array:4 [
            "nombre" => "M&#46;J&#46;"
            "apellidos" => "de los Santos"
            "email" => array:1 [
              0 => "mariajose.delossantos@ivirma.com"
            ]
            "referencia" => array:2 [
              0 => array:2 [
                "etiqueta" => "<span class="elsevierStyleSup">a</span>"
                "identificador" => "af0005"
              ]
              1 => array:2 [
                "etiqueta" => "<span class="elsevierStyleSup">&#42;</span>"
                "identificador" => "cr0005"
              ]
            ]
          ]
        ]
        "afiliaciones" => array:2 [
          0 => array:3 [
            "entidad" => "Laboratorio FIV&#44; Instituto Valenciano de Infertilidad&#44; IVI-RMA&#44; Valencia&#44; Spain"
            "etiqueta" => "a"
            "identificador" => "af0005"
          ]
          1 => array:3 [
            "entidad" => "Fundaci&#243;n IVI&#44; Valencia&#44; Spain"
            "etiqueta" => "b"
            "identificador" => "af0010"
          ]
        ]
        "correspondencia" => array:1 [
          0 => array:3 [
            "identificador" => "cr0005"
            "etiqueta" => "&#8270;"
            "correspondencia" => "Corresponding author at&#58; Laboratorio de FIV&#44; Instituto Valenciano de Infertilidad&#44; IVI-RMA&#44; Pl&#46; de la Polic&#237;a Local&#44; 3&#44; Valencia 46015&#44; Spain&#46;"
          ]
        ]
      ]
    ]
    "titulosAlternativos" => array:1 [
      "es" => array:1 [
        "titulo" => "Cultivo embrionario extendido hasta 14 d&#237;as"
      ]
    ]
    "resumenGrafico" => array:2 [
      "original" => 0
      "multimedia" => array:8 [
        "identificador" => "f0010"
        "etiqueta" => "Figure 2"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr2.jpeg"
            "Alto" => 674
            "Ancho" => 1778
            "Tamanyo" => 119460
          ]
        ]
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "al0010"
            "detalle" => "Figure "
            "rol" => "short"
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="sp0010" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleBold">Events of human implantation morphogenesis&#46;</span> The embryo formed by the ICM &#40;inner cell mass&#41; and the trophectoderm is shown first&#46; On the one hand&#44; from the ICM&#44; the epiblast and hypoblast segregate&#44; forming the bilaminar disc&#46; In the epiblast&#44; a cavity called the pro-amniotic cavity begins to appear&#44; which will later give rise to the amniotic cavity&#44; and the hypoblast extends to form the yolk sac&#46; On the other hand&#44; the trophectoderm differentiates into cytotrophoblast and syncytiotrophoblast&#46;</p>"
        ]
      ]
    ]
    "textoCompleto" => "<span class="elsevierStyleSections"><span id="s0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0070">Introduction</span><p id="p0005" class="elsevierStylePara elsevierViewall">Human early embryonic development from long before pre-implantation to gastrulation has always been a very active but difficult field of developmental biology&#46;</p><p id="p0010" class="elsevierStylePara elsevierViewall">It begins with a series of morphogenetic divisions and rearrangements leading to the formation of the blastocyst&#44; constituted by the inner cell mass &#40;ICM&#41; and the trophectoderm&#44; which floats freely between days 5 or 6 of development&#46; Around the seventh day&#44; this human embryo&#44; now consisting of epiblast and primitive endoderm derived from the ICM and trophectoderm&#44; must implant in the maternal uterus to survive &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41;&#46; Implantation of the blastocyst is a milestone in early mammalian embryonic development&#44; as it is the moment at which a coordinated program of lineage diversification&#44; cell fate specification and morphogenetic movements establishes the generation of extraembryonic tissues and the embryo proper&#44; and determines the conditions for successful gestation and gastrulation &#40;<a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al&#46;&#44; 2016</a>&#41;&#46;</p><p id="p0015" class="elsevierStylePara elsevierViewall">Errors in this process can lead to adverse pregnancy outcomes&#44; including miscarriages and fetal defects&#44; so understanding the mechanisms underlying this early development is of great importance not only for basic developmental biology but also for regenerative medicine &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41; and for resolving many cases of infertility and early pregnancy loss &#40;<a class="elsevierStyleCrossRef" href="#bb0170">Lusine&#44; 2020</a>&#41;&#46;</p><p id="p0020" class="elsevierStylePara elsevierViewall">The first days of pre-implantation development have been extensively studied using methods pioneered in <span class="elsevierStyleItalic">in vitro</span> fertilization &#40;IVF&#41; &#40;<a class="elsevierStyleCrossRef" href="#bb0075">Edwards et al&#46;&#44; 1969</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0080">Edwards et al&#46;&#44; 1970</a>&#41;&#46; Most of our knowledge about human embryogenesis from pre-implantation to gastrulation derives from studies of existing anatomical and histological collections of human embryos &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41; and the majority of observations and experiments have been carried out using other mammals as models&#46; For example&#44; monkey embryos &#40;<a class="elsevierStyleCrossRef" href="#bb0085">Enders et al&#46;&#44; 1986</a>&#41; as well as <span class="elsevierStyleItalic">in vitro</span> implantation systems in mice &#40;<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al&#46;&#44; 2014</a>&#41; have been used as models for human embryo development&#46;</p><p id="p0025" class="elsevierStylePara elsevierViewall">Recent advances in the <span class="elsevierStyleItalic">in vitro</span> embryogenesis&#44; including the development of extended <span class="elsevierStyleItalic">in vitro</span> culture systems for mouse and primate embryos and the construction of stem cell-based embryo models&#44; with the help of multi-omics studies and single cell-based imaging techniques&#44; open new doors to study human embryonic development and have significantly improved our understanding of the characteristics and mechanisms of human embryos from pre-implantation to gastrulation &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41;&#46; However&#44; although embryology presents a similar general program in all mammals&#44; it should be noted that the extrapolation of data to humans is limited&#44; because mammalian embryos show species-specific differences in post-implantation morphology &#40;<a class="elsevierStyleCrossRef" href="#bb0240">Rossant&#44; 2015</a>&#41;&#46;</p><p id="p0030" class="elsevierStylePara elsevierViewall">Recently&#44; in order to explain what happens in the early stages after human embryo implantation&#44; the groups of <a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al&#46; &#40;2016&#41;</a> and <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46; &#40;2016&#41;</a> have adapted a protocol for <span class="elsevierStyleItalic">in vitro</span> culture of mouse embryos&#44; allowing human embryos to develop up to the maximum number of days legally allowed&#46;</p><p id="p0035" class="elsevierStylePara elsevierViewall">In the present work we aim to review aspects related culture conditions&#44; morphogenesis events&#44; potential applications of long-term <span class="elsevierStyleItalic">in vitro</span> culture of human embryos as well as some ethical issues surrounding this topic&#46;</p></span><span id="s0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0075">Materials and methods</span><p id="p0040" class="elsevierStylePara elsevierViewall">In the following study&#44; an electronic bibliographic review of several papers and publications obtained from different databases&#44; including Pubmed and the National Center for Biotechnology Information &#40;NCBI&#41;&#44; has been carried out&#46; In addition&#44; the academic search engine &#34;Google Scholar&#34; and different websites&#44; such as ThermoFisher Scientific&#44; have been used for some components of the culture medium&#46; The research of information took place between the end of 2021 and the beginning of 2022&#44; and the main materials used were papers in English or Spanish from the last five years&#46; The keywords used for the research of information were&#58; human embryo development&#44; <span class="elsevierStyleItalic">in vitro</span> embryo development&#44; pre-implantation&#44; <span class="elsevierStyleItalic">in vitro</span> embryo culture&#44; and 14-day rule&#46;</p></span><span id="s0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0080">Results and discussion</span><span id="s0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0085">What is extended embryo culture up to 14 days and what are its conditions&#63;</span><p id="p0045" class="elsevierStylePara elsevierViewall">This extended culture consists of culturing fresh or thawed embryos that are at day 5 or 6 of development up to the maximum of 14 days allowed by law&#46; But how has it been possible to culture human embryos <span class="elsevierStyleItalic">in vitro</span> up to this limit&#63;</p><p id="p0050" class="elsevierStylePara elsevierViewall">Before answering this question&#44; it should be noted that&#44; although long-term <span class="elsevierStyleItalic">in vitro</span> culture of mouse and primate embryos is promising&#44; the current conditions need to be optimised in terms of reproducibility and efficiency&#46; High-quality embryos and optimised cell culture environments &#40;culture medium&#44; supplements&#44; and gas regulation mechanism&#41; are essential for stable and efficient protocols for long-term mammalian embryo culture&#46; Furthermore&#44; in current long-term cultures&#44; extra-embryonic tissues tend to grow vigorously with embryo development&#44; hindering the development of the inner embryo by absorbing nutrients from the culture medium&#46; In this sense&#44; strategies to equilibrate extra-embryonic and embryonic growth may also be a good starting point to achieve long-term embryo culture&#44; as human embryo development is highly dependent on the simultaneous growth of extra-embryonic tissues that provide essential signals to help extend embryo development to certain stages &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41;&#46;</p><p id="p0055" class="elsevierStylePara elsevierViewall">Meanwhile&#44; the group of <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al&#46; &#40;2016&#41;</a> established a culture system that allowed human embryos to develop <span class="elsevierStyleItalic">in vitro</span> through post-implantation stages&#46; This was based on a system for culturing mouse embryos through implantation &#40;<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al&#46;&#44; 2014</a>&#41;&#46;</p><p id="p0060" class="elsevierStylePara elsevierViewall">This procedure basically consists of thawing the embryos &#40;if vitrified&#41;&#44; removing the zona pellucida with tyrodes acid&#44; and placing them in culture until day 14 &#40;<a class="elsevierStyleCrossRef" href="#f0005">figure 1</a>&#41;&#46; Interestingly&#44; as in the sequential approach used in IVF to culture embryos from day 3 to blastocyst culture to provide the embryos with a specific nutrient environment&#44; long-term culture requires a gradual replacement of culture media containing decreasing amounts of serum&#46; The first 48 hours of culture that is from day 5-6 to day 7-8&#44; embryos are cultured using first culture medium &#40;IVC1&#41;&#46; On day 9-10&#44; half of the culture medium is replaced by another medium &#40;IVC2&#41; and renewed daily until day 14 by removing half of the medium and adding the other half&#46; More detailed information can be found in <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46; &#40;2016&#41;</a> and in <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al&#46; &#40;2016&#41;</a>&#46;</p><elsevierMultimedia ident="f0005"></elsevierMultimedia></span><span id="s0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0090">The conditions and components of the extended culture medium are explained below</span><p id="p0065" class="elsevierStylePara elsevierViewall">Like conventional embryo culture&#44; long-term culture has&#44; with some particularities&#44; similar culture conditions requirements to conventional embryo culture in IVF&#46;</p><span id="s0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0095">Temperature</span><p id="p0070" class="elsevierStylePara elsevierViewall">With regard to the optimal culture temperature&#44; it depends on the body temperature of the animal from which the cells are collected and the regional temperature variation in that animal&#46; Some studies have shown that the temperature within the female reproductive tract can be lower than 37&#176;C&#44; indicating that a lower temperature may be more beneficial for embryo development &#40;<a class="elsevierStyleCrossRef" href="#bb0015">Baak et al&#46;&#44; 2019</a>&#41;&#46; Indeed successful fertilization&#44; blastocyst formation rates and pregnancies can be achieved at 36&#176;C &#40;<a class="elsevierStyleCrossRef" href="#bb0125">Hong et al&#46;&#44; 2014</a>&#41;&#46;</p><p id="p0075" class="elsevierStylePara elsevierViewall">Currently&#44; the temperature used in the long-term culture is also 37&#176;C &#40;<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al&#46;&#44; 2014</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al&#46;&#44; 2019</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0260">Shahbazi et al&#46;&#44; 2020</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0340">Weimar et al&#46;&#44; 2013</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al&#46;&#44; 2016</a>&#41;&#46;</p></span><span id="s0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0100">Gas environment</span><p id="p0080" class="elsevierStylePara elsevierViewall">Carbon dioxide &#40;CO<span class="elsevierStyleInf">2</span>&#41; and oxygen &#40;O<span class="elsevierStyleInf">2</span>&#41; levels inside the culture incubator must also be controlled&#46; CO<span class="elsevierStyleInf">2</span> is essential to maintain the right pH in the culture medium&#44; which ranges between 7&#46;2-7&#46;4&#44; and allow proper embryo development &#40;<a class="elsevierStyleCrossRef" href="#bb0225">Reimundo et al&#46;&#44; 2021</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0230">Rend&#243;n Abad&#44; 2018</a>&#41;&#46; As in IVF&#44; 5&#37; CO<span class="elsevierStyleInf">2</span> concentration is commonly used for embryo culture up to day 14 &#40;<a class="elsevierStyleCrossRef" href="#bb0025">Bedzhov et al&#46;&#44; 2014</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al&#46;&#44; 2019</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0260">Shahbazi et al&#46;&#44; 2020</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0340">Weimar et al&#46;&#44; 2013</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al&#46;&#44; 2016</a>&#41;&#46;</p><p id="p0085" class="elsevierStylePara elsevierViewall">Regarding O<span class="elsevierStyleInf">2</span> concentration&#44; the trend in IVF clinics over the past years has been to culture pre-implantation human embryos in hypoxic conditions &#40;5&#37; O<span class="elsevierStyleInf">2</span>&#41; &#40;<a class="elsevierStyleCrossRef" href="#bb0060">Christianson et al&#46;&#44; 2014</a>&#41;&#46; Low O<span class="elsevierStyleInf">2</span> tension improves both embryo development and IVF outcomes &#40;<a class="elsevierStyleCrossRef" href="#bb0065">De los Santos et al&#46;&#44; 2013</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0320">van Montfoort et al&#46;&#44; 2020</a>&#41;&#46; When data from the meta-analyses were stratified for duration of culture under low O<span class="elsevierStyleInf">2</span>&#44; it was demonstrated that even though embryo quality was lower at cleavage stage &#40;<a class="elsevierStyleCrossRef" href="#bb0065">De los Santos et al&#46;&#44; 2013</a>&#41;&#44; pregnancy or live birth rates were only higher when the blastocyst stage &#40;<a class="elsevierStyleCrossRef" href="#bb0190">Nastri et al&#46;&#44; 2016</a>&#41;&#46;</p><p id="p0090" class="elsevierStylePara elsevierViewall">As the embryo is more vulnerable to oxygen in the pre-compaction stage as compared to the post-compaction stage&#44; low O<span class="elsevierStyleInf">2</span> is recommended from the initial stages of development &#40;<a class="elsevierStyleCrossRef" href="#bb0325">Wale and Gardner&#44; 2010</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0165">Kirkegaard et al&#46;&#44; 2013</a>&#41;&#46; However&#44; it has been observed that beyond day 7 of development&#44; embryos may benefit from higher O<span class="elsevierStyleInf">2</span> concentrations &#40;21&#37;&#41;&#44; possibly due to the increased size of the embryo&#44; which may decrease the O<span class="elsevierStyleInf">2</span> pressure in the embryonic nucleus&#46; Hypoxia clearly has a complex influence on embryonic development and its effects need to be better understood in the intrauterine environment to determine exactly how it affects embryogenesis &#40;<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#41;&#46; With regard to the concentration of this gas&#44; although some authors have used hypoxia conditions &#40;<a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al&#46;&#44; 2019</a>&#41;&#44; most of them have done the extended culture until day 14 with an O<span class="elsevierStyleInf">2</span> concentration of 21&#37; &#40;<a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0260">Shahbazi et al&#46;&#44; 2020</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al&#46;&#44; 2016</a>&#41;&#46;</p></span><span id="s0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0105">Culture media composition</span><p id="p0095" class="elsevierStylePara elsevierViewall">The composition of the long-term culture medium is slightly modified compared to the embryo culture media widely used in IVF laboratories such as G-2<span class="elsevierStyleSup">TM</span> &#40;Vitrolife&#44; Sweden&#41; or Sydney IVF Blastocyst Medium &#40;Cook Medical&#44; United States&#41; as they do not contain hormones&#44; nor serum from animal sources&#46;</p><p id="p0100" class="elsevierStylePara elsevierViewall">Indeed&#44; both IVC1 and IVC2&#44; are made up of the same components such as advanced DMEM&#47;F12&#44; ITS-X&#44; L-glutamine&#44; &#946;-oestradiol&#44; progesterone&#44; penicillin&#47;streptomycin&#44; and N-acetyl-L-cysteine&#46; Whereas IVC1 is supplemented with 20&#37; heat-inactivated FBS&#44; IVC2 by 30&#37; Knockout Serum Replacement&#46;</p><p id="p0105" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleBold">Advanced DMEM&#47;F12</span> &#40;ThermoFisher Scientific&#44; United States&#41; which used as basal medium&#44; is composed&#44; like the conventional IVF media mentioned above&#44; of amino acids&#44; inorganic salts&#44; vitamins&#44; proteins&#44; and glucose&#46; However&#44; it has certain peculiarities&#46; It does not contain human serum albumin &#40;HSA&#41;&#44; but instead includes other proteins such as AlbuMAX&#174; II &#40;Gibco&#44; United States&#41;&#44; human transferrin and recombinant full-chain insulin&#46; Furthermore&#44; it contains additional components such as fatty acids like linoleic acid and linolenic acid&#44; reducing agents like glutathione&#44; monosodium&#44; and other components like putrescine or thymidine &#40;all components and concentrations are specified in <a class="elsevierStyleCrossRef" href="#t0005">Table 1</a>&#41; &#40;<a class="elsevierStyleCrossRef" href="#bb0300">ThermoFisher Scientific&#44; 2020a</a>&#41;&#46; Despite this&#44; the <span class="elsevierStyleItalic">in vitro</span> function of some of these components on the embryo has not been completely described&#46;</p><elsevierMultimedia ident="t0005"></elsevierMultimedia><p id="p0110" class="elsevierStylePara elsevierViewall">On the other hand <span class="elsevierStyleBold">ITS-X</span>&#44; which is composed of&#58; &#40;1&#41; insulin&#44; which promotes glucose and amino acid uptake&#44; lipogenesis&#44; intracellular transport&#44; and protein and nucleic acid synthesis&#59; &#40;2&#41; transferrin&#44; which acts as an iron carrier and can also help reduce toxic levels of oxygen radicals and peroxide&#59; &#40;3&#41; selenium&#44; as sodium selenite&#44; which acts as a cofactor for glutathione peroxidase and other proteins that is used as an antioxidant in the medium&#59; and &#40;4&#41; ethanolamine&#44; a precursor of phosphoglycerides that are essential for the structure of the plasma membrane and cellular organelles &#40;Concentrations are specified in <a class="elsevierStyleCrossRef" href="#t0005">Table 1</a>&#41;&#46; All these components have been shown to reduce the need for serum supplementation of conventional cell culture media and to promote optimal performance of serum-free media &#40;<a class="elsevierStyleCrossRef" href="#bb0290">ThermoFisher Scientific&#44; 2014b</a>&#41;&#46; This is the case for the culture media up to day 14&#44; IVC1 and IVC2 respectively&#44; where IVC1 contains only 20&#37; inactivated FBS and IVC2 contains Knockout Serum Replacement&#44; which replaces FBS&#46; Possible reasons for using reduced serum reduced serum are unknown exact composition&#44; batch-to-batch variability&#44; unintended interaction with culture substances&#44; potentially leading to unexpected or undesired results&#44; experimental variability&#44; and limited reproducibility &#40;<a class="elsevierStyleCrossRef" href="#bb0315">van der Valk et al&#46;&#44; 2018</a>&#41;&#46; In addition&#44; with Knockout Serum Replacement&#44; many of the disadvantages of using FBS in culture can be mitigated &#40;<a class="elsevierStyleCrossRef" href="#bb0300">ThermoFisher Scientific&#44; 2020a</a>&#41;&#46;</p><p id="p0115" class="elsevierStylePara elsevierViewall">As discussed&#44; both conventional and long-term culture media contain <span class="elsevierStyleBold">amino acids</span> in their composition&#46; These molecules have numerous functions&#44; and their importance is attributed to acting as precursors for protein and nucleotide biosynthesis&#44; substrates for energy metabolism&#44; osmolytes&#44; antioxidant&#44; regulators&#44; buffers&#44; and chelators&#46; From studies in mice&#44; non-essential amino acids stimulate the rate of cleavage&#44; blastocyst formation and hatching&#46; Essential amino acids stimulate cleavage rates after the eight-cell stage and increase the development of the inner cell mass in the blastocyst &#40;<a class="elsevierStyleCrossRef" href="#bb0280">Sunde et al&#46;&#44; 2016</a>&#41;&#46;</p><p id="p0120" class="elsevierStylePara elsevierViewall">As in IVF culture media&#44; L-glutamine is not present&#44; instead <span class="elsevierStyleBold">GlutaMAX</span><span class="elsevierStyleSup"><span class="elsevierStyleBold">TM</span></span> &#40;ThermoFisher Scientific&#44; United Stated&#41; is added as an alternative&#46; This supplement consists of the dipeptide L-alanyl-L-glutamine&#44; which is more stable in aqueous solutions&#44; making it suitable for suspension and adherent cultures of mammalian cells&#44; such as embryos cultured up to day 14&#46; Moreover&#44; compared to L-glutamine&#44; GlutaMAX minimises toxic ammonia accumulation&#44; improves cell viability and growth by having lower ammonia levels&#44; and remains stable over a wide temperature range because it does not spontaneously degrade &#40;<a class="elsevierStyleCrossRef" href="#bb0305">ThermoFisher Scientific&#44; 2020b</a>&#41;&#46; It should be noted that&#44; although this supplement as such is not added to conventional culture media used in IVF laboratories&#44; they contain stabilised glutamine in the form of L-alanine-L-glutamine&#46;</p><p id="p0125" class="elsevierStylePara elsevierViewall">Another peculiarity of the basal medium of the long-term culture is that it contains a compound called <span class="elsevierStyleBold">AlbuMAX&#174; II</span>&#46; Conventional culture medium contains HSA with associated lipids&#46; However&#44; this supplement consists of lipid-rich bovine serum albumin &#40;BSA&#41;&#44; and like ITS-X&#44; is effective in reducing or replacing the requirement for serum supplementation in culture media&#46; In addition&#44; the chromatographic separation process contributes to the preservation of the natural lipids associated with the purified albumin&#44; resulting in excellent growth stimulation characteristics &#40;<a class="elsevierStyleCrossRef" href="#bb0295">ThermoFisher Scientific&#44; 2014c</a>&#41;&#46;</p><p id="p0130" class="elsevierStylePara elsevierViewall">Additionally&#44; to these protein-associated lipids&#44; the long-term culture medium contains other <span class="elsevierStyleBold">lipids</span>&#44; specifically fatty acids&#44; which are not added normally in conventional IVF media such as those mentioned above&#46; It is known that embryos need lipids at very early stages&#46; Most of the ATP for pre-implantation development comes from fatty acids&#46; The proportion of different types of lipids plays an important role&#46; An increase in unsaturated fatty acids can compensate for the negative effects of saturated fatty acids&#46; Polyunsaturated fatty acids &#40;linoleic acid and linolenic acid&#41; have a beneficial role in embryonic cells signalling and may influence gene expression in lipid metabolism&#44; oxidation and fatty acid synthesis as seen in animal models &#40;<a class="elsevierStyleCrossRef" href="#bb0210">Pawar and Jump&#44; 2003</a>&#41;&#46; The impact of trans-unsaturated fatty acids on embryonic development is unknown&#46; However&#44; it is thought that their effect on polyunsaturated fatty acid metabolism could affect embryonic development &#40;<a class="elsevierStyleCrossRef" href="#bb0095">F&#252;rstova et al&#46;&#44; 2008</a>&#41;&#46; This means that lipid balance may be important for pre-implantation embryo development and implantation &#40;<a class="elsevierStyleCrossRef" href="#bb0355">Zander-Fox et al&#46;&#44; 2021</a>&#41;&#46;</p><p id="p0135" class="elsevierStylePara elsevierViewall">The basal medium also contains&#44; although at low concentration&#44; polyamines such as <span class="elsevierStyleBold">putrescine</span>&#46; These play a key role in cell proliferation&#44; growth&#44; and differentiation&#46; They are involved in the regulation of DNA and protein synthesis&#44; proliferation and differentiation &#40;<a class="elsevierStyleCrossRef" href="#bb0160">Igarashi and Kashiwagi&#44; 2000</a>&#41;&#46; They act as antioxidants protecting DNA&#44; proteins and lipids from oxidative damage &#40;<a class="elsevierStyleCrossRef" href="#bb0055">Chattopadhyay et al&#46;&#44; 2003</a>&#41;&#46; There are evidences that these molecules regulate early embryogenesis&#44; placental trophoblastic growth&#44; implantation and embryonic development &#40;<a class="elsevierStyleCrossRef" href="#bb0110">Halloran et al&#46;&#44; 2021</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0350">Wu and Morris Jr&#44; 1998</a>&#41;&#46; In addition&#44; supplementation of polyamines such as putrescine has been shown to be involved in embryo survival&#44; ultimately resulting in improved fertility &#40;<a class="elsevierStyleCrossRef" href="#bb0145">Hussain et al&#46;&#44; 2017</a>&#41;&#46;</p><p id="p0140" class="elsevierStylePara elsevierViewall">Another component present in the culture media is <span class="elsevierStyleBold">vitamins</span>&#46; In contrast to conventional media&#44; the culture up to day 14 contains an abundance of vitamins B and C&#46; Although the role of some of them in embryonic development is known&#44; the role of others is unknown&#46; In hamsters&#44; vitamins B have been shown to promote embryonic development <span class="elsevierStyleItalic">in vitro</span>&#59; however&#44; they may be inhibitory in early mouse embryos&#46; Vitamins B2 &#40;riboflavin&#41;&#44; 6 &#40;pyridoxine&#41;&#44; 9 &#40;folic acid&#41;&#44; 12 &#40;cobalamin&#41; are known to be necessary for homocysteine recycling &#40;<a class="elsevierStyleCrossRef" href="#bb0185">M&#233;n&#233;zo et al&#46;&#44; 2013</a>&#41;&#46; Vitamin C &#40;ascorbic acid&#41; acts as an antioxidant by reducing oxidative stress-induced embryo toxicity&#44; and in mice it also improves blastocyst development rates &#40;<a class="elsevierStyleCrossRef" href="#bb0330">Wang et al&#46;&#44; 2002</a>&#41;&#46;</p><p id="p0145" class="elsevierStylePara elsevierViewall">In addition to the mentioned molecules that act as antioxidants &#40;amino acids&#44; proteins&#44; vitamins&#41;&#44; the basal culture medium of the extended culture contains <span class="elsevierStyleBold">N-acetyl-L-cysteine</span> and <span class="elsevierStyleBold">reducing agents</span> such as glutathione&#44; monosodium which are not present in conventional IVF media&#46;</p><p id="p0150" class="elsevierStylePara elsevierViewall">As mentioned before&#44; the culture medium up to day 14 also contains <span class="elsevierStyleBold">hormones</span> which are not present in the routine culture media used in IVF laboratories&#46; There is a wealth of evidence that growth factors and hormones in the reproductive tract and in the pre-embryo may be potential regulators of embryonic development&#46; Their benefits are thought to include decreased apoptosis&#44; increased cell proliferation&#44; and consequently improved progression to blastocyst&#46; The need for their presence and usefulness in culture media is unclear due to the unknown long-term effects they may have on pre-embryos&#46; However&#44; culture media containing different growth factors and hormones can be found on the market &#40;<a class="elsevierStyleCrossRef" href="#bb0230">Rend&#243;n Abad&#44; 2018</a>&#41;&#46; Long-term culture medium&#44; in its initial protocol and unlike conventional IVF media&#44; contains progesterone and &#946;-oestradiol&#46;</p><p id="p0155" class="elsevierStylePara elsevierViewall">Extended culture medium will also include other supplements such as <span class="elsevierStyleBold">antibiotics</span> &#40;e&#46;g&#46; penicillin and streptomycin&#41;&#46; These are added to the medium to prevent or reduce likelihood of contamination&#46; Although gentamicin has been found to be the antibiotic used par excellence in culture media as a prophylactic measure&#44; it should be noted that the absence of antibiotics is beneficial for the development of pre-embryos as they develop faster &#40;<a class="elsevierStyleCrossRef" href="#bb0230">Rend&#243;n Abad&#44; 2018</a>&#41;&#46;</p></span><span id="s0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0110">What structures can we see with the culture up to day 14&#63;</span><p id="p0160" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">In vitro</span> culture systems up to day 14 of embryo development have made it possible to document the key events of normal human embryogenesis from pre-implantation to gastrulation&#46; By comparing embryos developing <span class="elsevierStyleItalic">in vitro</span> with the Carnegie series of human embryos <span class="elsevierStyleItalic">in vivo</span> &#40;<a class="elsevierStyleCrossRef" href="#bb0120">Hertig et al&#46;&#44; 1956</a>&#41;&#44; the main morphogenetic events of human implantation morphogenesis were discovered &#40;<a class="elsevierStyleCrossRef" href="#f0010">figure 2</a>&#41;&#46; These include&#58;<ul class="elsevierStyleList" id="l0005"><li class="elsevierStyleListItem" id="li0005"><span class="elsevierStyleLabel">1&#46;</span><p id="p0165" class="elsevierStylePara elsevierViewall">Segregation of the epiblast and hypoblast from the inner cell mass of the blastocyst and morphogenetic movements to form the bilaminar disc&#46; While the epiblast is represented by increased levels of OCT4 and Nanog &#40;pluripotency transcription factors&#41;&#44; the hypoblast is determined by increased levels of GATA6 and Sox17 &#40;hypoblast-specific transcription factors&#41;&#46;</p></li><li class="elsevierStyleListItem" id="li0010"><span class="elsevierStyleLabel">2&#46;</span><p id="p0170" class="elsevierStylePara elsevierViewall">Pro-amniotic cavity formation within pluripotent epiblast cells of the embryonic lineage &#40;via apically located atypical PKC&#44; leading to apical polarity in epiblast cells and lumen formation&#41; and amniotic epithelium&#46;</p></li><li class="elsevierStyleListItem" id="li0015"><span class="elsevierStyleLabel">3&#46;</span><p id="p0175" class="elsevierStylePara elsevierViewall">Formation of the embryonic disc from the epiblast &#40;embryo proper&#41;&#46;</p></li><li class="elsevierStyleListItem" id="li0020"><span class="elsevierStyleLabel">4&#46;</span><p id="p0180" class="elsevierStylePara elsevierViewall">Formation of the primary yolk sac within the hypoblast&#46;</p></li><li class="elsevierStyleListItem" id="li0025"><span class="elsevierStyleLabel">5&#46;</span><p id="p0185" class="elsevierStylePara elsevierViewall">Differentiation of the trophoblast to produce its two characteristic cell types&#58; the mononucleated cytotrophoblast and the multinucleated polyploid syncytiotrophoblast&#46;</p></li></ul></p><elsevierMultimedia ident="f0010"></elsevierMultimedia><p id="p0190" class="elsevierStylePara elsevierViewall">Despite not all embryos will be able to undergo the former morphogenic event&#44; the system brings <span class="elsevierStyleItalic">in vitro</span> cultured embryos to the point of gastrulation &#40;<a class="elsevierStyleCrossRef" href="#bb0250">Shahbazi and Zernicka-Goetz&#44; 2018</a>&#41;&#46; Interestingly&#44; all of them can take place <span class="elsevierStyleItalic">in vitro</span> in the absence of maternal tissues&#44; indicating that human embryos have a previously underestimated potential for self-organization &#40;<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0360">Zernicka-Goetz et al&#46;&#44; 2016</a>&#41;&#46;</p><p id="p0195" class="elsevierStylePara elsevierViewall">Although this system may not be able to fully recapitulate all aspects of human embryogenesis <span class="elsevierStyleItalic">in vivo</span>&#44; it has revealed that even in the absence of endometrial cells&#44; human blastocysts have remarkable capacity for self-organization that was previously unknown &#40;<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#41;&#46;</p></span><span id="s0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0115">Stem cells and culture</span><p id="p0200" class="elsevierStylePara elsevierViewall">In recent years&#44; the cultivation of stem cell-based embryonic models to unravel the mysteries of early embryonic development in humans has gained ground&#46;</p><p id="p0205" class="elsevierStylePara elsevierViewall">The stem cell-based embryo model is a simplified model that attempts to mimic the characteristics of a natural embryo&#46; The knowledge gained from their experimentation opens a new way to model embryogenesis&#44; cell lineage differentiation&#44; tissue morphogenesis&#44; and organogenesis in mammalian development &#40;<a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam&#44; 2021</a>&#41;&#46; Moreover&#44; it can improve the understanding of the mechanisms of embryo development from blastocyst stage to gastrulation&#44; as it is more scalable&#44; versatile&#44; accessible than natural embryos&#44; and exhibits similar self-organization as natural embryos during <span class="elsevierStyleItalic">in vitro</span> culture &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41;&#46;</p><p id="p0210" class="elsevierStylePara elsevierViewall">There are two types of stem cell-based models &#40;<a class="elsevierStyleCrossRef" href="#f0015">figure 3</a>&#41;&#46; On the one hand&#44; <span class="elsevierStyleItalic">non-integrated models</span> use a single stem cell type &#40;ESC&#41; and are limited to the formation of specific parts of the embryo such as amniotic sac structures and shed light on selected morphogenetic events such as gastrulation or developmental processes such as germ layer differentiation&#46; This type of model includes amniotic sac models &#40;<a class="elsevierStyleCrossRef" href="#bb0370">Zheng et al&#46;&#44; 2019</a>&#41; and the elongated gastruloid resembling the posterior elongation body axis &#40;<a class="elsevierStyleCrossRef" href="#bb0020">Beccari et al&#46;&#44; 2018</a>&#41;&#46; On the other hand&#44; <span class="elsevierStyleItalic">integrated models</span> are formed from several stem cell types &#40;ESC&#44; XENC&#44; TSC&#41; and contain all parts of the conceptus&#44; that is&#44; embryonic and extra-embryonic tissues&#44; giving rise to the blastoid&#44; a blastocyst-like structure formed by the aggregate of ESC and TSC&#44; where TSC encloses ESC in a cavity &#40;<a class="elsevierStyleCrossRef" href="#bb0235">Rivron et al&#46;&#44; 2018</a>&#41; and the embryoid&#44; an embryo-like structure made up of various types of stem cells but not capable of developing into a functional human embryo &#40;<a class="elsevierStyleCrossRef" href="#bb0270">Sozen et al&#46;&#44; 2018</a>&#41;&#46; Both types of models can be achieved by means of 2D micropatterning techniques&#44; microfluidic devices&#44; or 3D aggregation methods&#44; exhibiting a self-organization similar to that of natural embryos during <span class="elsevierStyleItalic">in vitro</span> culture&#46; It should be noted that most of this type of work has now been carried out in animal models such as the mouse &#40;<a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam&#44; 2021</a>&#41;&#46;</p><elsevierMultimedia ident="f0015"></elsevierMultimedia><p id="p0215" class="elsevierStylePara elsevierViewall">Limited knowledge about the mechanisms regulating cell fate determination during early human embryogenesis hampers the creation of stem cell-based embryo models&#46; This could only be remedied by further research with human embryos per se&#44; non-human primate embryos <span class="elsevierStyleItalic">in vitro</span> and embryo models &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41;&#46; And&#44; although these culture systems could reproduce the events of early mammalian embryogenesis <span class="elsevierStyleItalic">in vivo</span>&#44; a critical prerequisite of experimental models is that they must accurately reconstruct the events that occur <span class="elsevierStyleItalic">in vivo</span>&#44; particularly in the early stages when the basic body plan is established and when the <span class="elsevierStyleItalic">in vivo</span> embryo is most inaccessible for experimentation &#40;<a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam&#44; 2021</a>&#41;&#46; By collecting more data on this&#44; extended <span class="elsevierStyleItalic">ex vivo</span> phenotype&#44; new markers of embryo competence and sustained implantation could be identified to enhance IVF outcomes&#46;</p></span><span id="s0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0120">Potential applications in embryology</span><p id="p0220" class="elsevierStylePara elsevierViewall">The contributions of post-implantation embryo research&#44; apart from allowing to observe <span class="elsevierStyleItalic">in vitro</span> the events and structures mentioned above&#44; could include advancing the understanding of gene function during embryogenesis&#59; a scientific basis for the prevention of early pregnancy loss&#44; birth defects and teratogenesis&#59; and an understanding of how the widespread epigenetic programming that occurs during this stage of development might affect disease progression in later life &#40;<a class="elsevierStyleCrossRef" href="#bb0215">Pera&#44; 2017</a>&#41;&#46;</p><p id="p0225" class="elsevierStylePara elsevierViewall">Advances in <span class="elsevierStyleItalic">in vitro</span> culture systems have allowed the modelling of chromosomal instability and the investigation of how mosaicism and chromosomal abnormalities can affect development in the pre-implantation period&#46;</p><p id="p0230" class="elsevierStylePara elsevierViewall">Before post-implantation <span class="elsevierStyleItalic">in vitro</span> culture&#44; the capacity of mosaic blastocysts had only been studied in mice &#40;<a class="elsevierStyleCrossRef" href="#bb0035">Bolton et al&#46;&#44; 2016</a>&#41; or through clinical outcome data &#40;<a class="elsevierStyleCrossRef" href="#bb0100">Greco et al&#46;&#44; 2015</a>&#41; with no blinded studies evaluating the IVF outcome of the reported mosaicism&#46; Without knowing the implications of mosaic embryos in IVF transfers&#44; it is difficult to determine whether to discard them and waste a potentially viable embryo or transfer a potentially harmful one&#46; <a class="elsevierStyleCrossRef" href="#bb0220">Popovic et al&#46; &#40;2019&#41;</a>&#44; using NGS and extended culture to day 12 of development&#44; are believed to be the first to investigate the fate of chromosomal abnormalities and mosaicism&#46; On the one hand&#44; they showed that many mosaics in the day 5 or 6 blastocyst biopsy were no longer mosaic at 8 or 12 days&#44; others were still mosaic&#44; and others had become uniformly aneuploid&#46; In general&#44; when mosaic blastocysts had more abnormal cells&#44; they were more likely to be non-viable at 12 days&#46; On the other hand&#44; they also showed that NGS can have an 18&#37; misdiagnosis rate for mosaicism&#44; so improvements and further studies are needed before clinical application&#46; However&#44; it shows the possible developments of these advances in extended culture &#40;<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson&#44; 2020</a>&#41;&#46;</p><p id="p0235" class="elsevierStylePara elsevierViewall">Consensus on the clinical management of embryonic mosaicism is still under debate&#46; <span class="elsevierStyleItalic">In vitro</span> performance of specific mosaics are needed to understand their developmental behaviour and improve diagnosis&#46; Furthermore&#44; assessing chromosomal instability during these hitherto hidden developmental stages may provide valuable information on the predictive value of reporting mosaicism in clinical practice&#44; ultimately improving the embryo selection process and clinical outcomes&#46;</p></span><span id="s0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0125">14-day limit</span><p id="p0240" class="elsevierStylePara elsevierViewall">The 14-day limit for <span class="elsevierStyleItalic">in vitro</span> human embryo culture dates back to the 1970s&#44; more specifically to 1978&#44; when the ability to develop embryos <span class="elsevierStyleItalic">in vitro</span> for research and assisted reproduction was demonstrated &#40;<a class="elsevierStyleCrossRef" href="#bb0275">Steptoe and Edwards&#44; 1978</a>&#41;&#46; But it was not until 1984 that it was first established in the Warnock Report in the UK &#40;<a class="elsevierStyleCrossRef" href="#bb0335">Warnock&#44; 1984</a>&#41;&#44; and subsequently enshrined in the <a class="elsevierStyleCrossRefs" href="#bb0130">Human Fertilization and Embryology &#40;HFE&#41; Acts of&#44; 1990&#44; 2008</a>&#46; It is now one of the most internationally accepted standards in reproductive medicine &#40;<a class="elsevierStyleCrossRef" href="#bb0010">Appleby and Bredenoord&#44; 2018</a>&#41;&#46;</p><p id="p0245" class="elsevierStylePara elsevierViewall">The Warnock Report &#40;1984&#41; elicited different responses from the public and committee members as there was a wide range of opinions on the moral status of human embryos&#46; However&#44; most of the committee agreed that such research should be allowed under licence&#46; To allay public fears it was necessary to set a clear limit&#44; which was 14 days&#44; that could be justified on the basis of this moral status of the embryo &#40;<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger&#44; 2021</a>&#41;&#46;</p><p id="p0250" class="elsevierStylePara elsevierViewall">One of the reasons for setting the maximum at 14 days was that this point in development was prior to the formation of the primitive line&#44; which appears on day 15 of human embryonic development&#46; This occurs at the onset of gastrulation&#44; a period when the three germ layers are formed and the axes and body form are established&#44; marking the point of individualisation &#40;<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson&#44; 2020</a>&#41;&#46; The committee&#39;s view was that from this point onwards the embryo could be considered as a defined individual and a potential person&#44; an entity with rights&#44; such as the right to life&#44; since from this point onwards twins cannot be formed&#46; Furthermore&#44; he pointed out that this boundary was prior to the formation of the central nervous system&#44; around day 22 of development&#44; which would exclude the possibility of the embryo experiencing pain &#40;<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger&#44; 2021</a>&#41;&#46; Other reasons were the substantial loss of embryos from fertilization to this point and that until the implantation process is not complete&#44; the embryo has no potential for further development &#40;<a class="elsevierStyleCrossRef" href="#bb0215">Pera&#44; 2017</a>&#41;&#46;</p><p id="p0255" class="elsevierStylePara elsevierViewall">However&#44; given that scientific progress has brought us to a point where viable pregnancies can be obtained from day 7 blastocyst transfer&#44; could be reasonable to question whether 14 days is too restrictive from a research point of view &#40;<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson&#44; 2020</a>&#41;&#46; Furthermore&#44; as discussed before&#44; recent studies have demonstrated the successful growth of human embryos up to developmental stages close to this limit &#40;<a class="elsevierStyleCrossRef" href="#bb0070">Deglincerti et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#41;&#46; Other research has reported the generation of embryo-like structures from mouse and human pluripotent stem cells that mimic the gastrulating embryo in form and cellular content &#40;<a class="elsevierStyleCrossRef" href="#bb0090">Etoc et al&#46;&#44; 2016</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0115">Harrison et al&#46;&#44; 2017</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0265">Shao et al&#46;&#44; 2017</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0310">van den Brink et al&#46;&#44; 2014</a>&#41;&#46; And recently&#44; successful <span class="elsevierStyleItalic">in vitro</span> culture of <span class="elsevierStyleItalic">Cynomolgus</span> macaque embryos beyond the gastrula stage and up to day 20 of development has been reported &#40;<a class="elsevierStyleCrossRef" href="#bb0195">Niu et al&#46;&#44; 2019</a>&#41;&#46; This may fill the knowledge gap on human embryogenesis beyond day 14&#44; and it is even suggested that such a culture period should also be feasible for human embryos &#40;<a class="elsevierStyleCrossRef" href="#bb0345">Williams and Johnson&#44; 2020</a>&#41;&#46;</p><p id="p0260" class="elsevierStylePara elsevierViewall">All these advances have led to a re-examination of the current restrictions on human embryo culture &#40;<a class="elsevierStyleCrossRef" href="#bb0150">Hyun et al&#46;&#44; 2016</a>&#41;&#44; with various positions or sides emerging&#46; There are those in favour of such an extension &#40;<a class="elsevierStyleCrossRef" href="#bb0155">Hyun et al&#46;&#44; 2021</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0175">McCully&#44; 2021</a>&#41;&#59; others who think that it should not be extended and should remain as it currently stands &#40;<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger&#44; 2021</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0105">Green et al&#46;&#44; 2021</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0180">McLaren&#44; 1984</a>&#41;&#59; and even the most extreme who argue that human embryos should not be experimented on&#46; Making changes to existing legislation is a complex process and there is a risk of creating new restrictions that could have unintended consequences for future research &#40;<a class="elsevierStyleCrossRef" href="#bb0215">Pera&#44; 2017</a>&#41;&#46;</p></span><span id="s0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0130">So&#44; should the 14-day limit be extended&#63;</span><p id="p0265" class="elsevierStylePara elsevierViewall">The period between days 14 and 28 of human embryonic development is called the &#34;black box&#34; &#40;<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al&#46;&#44; 2017</a>&#41;&#46; Until now&#44; embryonic development in this period has been difficult to study and understand from a scientific and regulatory point of view&#46; However&#44; the <a class="elsevierStyleCrossRef" href="#bb0200">Nuffield Council on Bioethics &#40;2017&#41;</a> argues that any attempt to extend research beyond day 14 would have to result in significant advances in science&#46; Furthermore&#44; it should examine the committee&#39;s reasoning and justify the extension on this basis&#46; The two key considerations are maintaining public confidence and providing a moral justification for embryo research&#46; The committee provided two reasons for morally justifying early embryo research&#58; the utilitarian argument&#44; based on the benefits of such research&#44; and its argument based on the moral status of early embryos &#40;<a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger&#44; 2021</a>&#41;&#46;</p><p id="p0270" class="elsevierStylePara elsevierViewall">Due to the current benefits of embryo research and the potential future benefits&#44; supporters of extending the 14-day rule are calling for its revision&#46; Extending this period to 28 days would allow to study the developmental processes of gastrulation&#44; when the first primitive tissues are formed&#46; In addition&#44; it would allow learning about the developing nervous system without the risk of neural connections and a better understanding of early organ development &#40;<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al&#46;&#44; 2017</a>&#41;&#46; It would also be possible to advance our understanding of cell fate decisions during early embryonic development &#40;<a class="elsevierStyleCrossRef" href="#bb0255">Shahbazi et al&#46;&#44; 2016</a>&#41;&#46; The safety and success of current IVF procedures could be further improved by understanding the nature of some birth defects and predicting which embryos are most likely to result in a successful pregnancy &#40;<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al&#46;&#44; 2017</a>&#41;&#46; Finally&#44; more could be learned about the physiology of pregnancy beyond day 14&#44; including the processes surrounding implantation&#44; a greater understanding of the causes of embryonic defects and why events such as miscarriages occur &#40;<a class="elsevierStyleCrossRef" href="#bb0140">Hurlbut et al&#46;&#44; 2017</a>&#41;&#46; <a class="elsevierStyleCrossRef" href="#bb0045">Castelyn &#40;2020&#41;</a> offers several reasons why a re-evaluation of the 14-day rule may be justified&#46;</p><p id="p0275" class="elsevierStylePara elsevierViewall">Sophia McCully takes this side&#44; listing several potential advantages to extending the limit to day 28 of development &#40;<a class="elsevierStyleCrossRef" href="#bb0175">McCully&#44; 2021</a>&#41;&#46; She focuses mainly on the utilitarian argument to justify her idea&#44; suggesting that it will help reduce miscarriage rates&#44; improve IVF outcomes&#44; and test the safety of new techniques such as gene editing&#46; And&#44; while acknowledging that the current limit has generated public confidence&#44; she argues that concerns about a slippery slope are unjustified because the regulations are strict&#44; making it illegal to slide down such a slope&#46; Other authors advocate a more conservative extension of about 2-3 days&#44; and that only after broad public consultation &#40;<a class="elsevierStyleCrossRef" href="#bb0155">Hyun et al&#46;&#44; 2021</a>&#41;&#46;</p><p id="p0280" class="elsevierStylePara elsevierViewall">However&#44; although it has important potential scientific and medical benefits&#44; it is unclear whether extending the 14-day limit in human embryo culture is the best way to achieve them or not&#44; as the possibility of prolonged normal development of human embryos <span class="elsevierStyleItalic">in vitro</span> during and beyond the primitive line stage is unknown&#46; In mice&#44; pre-implantation embryo culture has so far not generated structures that resemble post-implantation embryos&#44; in contrast to recent results with stem cell cultures&#46; In humans&#44; degenerative changes have occurred as they approach the 14-day limit with the current methodology&#46; In mice&#44; embryos have been grown <span class="elsevierStyleItalic">in vitro</span> up to day 10&#46;5 of development&#44; the equivalent of about 4 weeks in humans&#44; but this is obviously unethical in our species&#46; So&#44; the question is whether it is feasible to extend <span class="elsevierStyleItalic">in vitro</span> development of high-quality human embryos to this stage&#46; However&#44; even if it were&#44; the extremely limited supply of such embryos means that any line of research requiring large numbers of cells would be extremely difficult to conduct and replicate in multiple laboratories &#40;<a class="elsevierStyleCrossRef" href="#bb0215">Pera&#44; 2017</a>&#41;&#46;</p><p id="p0285" class="elsevierStylePara elsevierViewall"><a class="elsevierStyleCrossRef" href="#bb0105">Green et al&#46; &#40;2021&#41;</a>&#44; despite supporting human embryo research&#44; are not very receptive to the recommendations of the International Society for Stem Cell Research &#40;ISSCR&#41; to extend the 14-day limit&#44; as they consider that there are compelling reasons based on important biological events to maintain it&#44; and believe that this institution should offer more convincing arguments for abandoning the current limit&#46;</p><p id="p0290" class="elsevierStylePara elsevierViewall">Another group that is opposed to this extension is <a class="elsevierStyleCrossRef" href="#bb0030">Blackshaw and Rodger &#40;2021&#41;</a>&#46; In response to McCully&#44; they argue that the Warnock committee&#39;s reasons for the 14-day limit still apply despite advances in culturing human embryos <span class="elsevierStyleItalic">in vitro</span>&#46; These authors believe that McCully seems unaware that such an extension is precisely the kind of slippery slope that opponents of the original law warned about&#46; She also fails to take into account the argument based on the moral status of embryos&#58; that only after the primitive line appears is the embryo a distinct individual and potential person&#46; Moreover&#44; research on embryos between 7 and 14 days old has only just begun to be exploited and it seems rash to extend it to 28 days&#46; It is also not known how precisely post-implantation embryo culture would replicate the <span class="elsevierStyleItalic">in utero</span> environment&#44; which may limit its applicability beyond 14 days&#46;</p><p id="p0295" class="elsevierStylePara elsevierViewall">Therefore&#44; several practical obstacles must first be overcome to facilitate this extended study&#46; For example&#44; conditions must be devised and improved to keep an embryo sustained and alive in a suitable environment &#40;<a class="elsevierStyleCrossRef" href="#bb0005">Aach et al&#46;&#44; 2017</a>&#41;&#46; Although this challenge is difficult&#44; it could be overcome with the help of 3D bioprinting and organoids &#40;<a class="elsevierStyleCrossRef" href="#bb0005">Aach et al&#46;&#44; 2017</a>&#59; <a class="elsevierStyleCrossRef" href="#bb0040">Bredenoord et al&#46;&#44; 2017</a>&#41;&#46; To culture embryos at later developmental stages&#44; such as beyond day 10&#46;5 in mice or day 30 in macaques&#44; it is essential to establish a functional placenta and a functional vascularised umbilical cord&#46; This has been achieved artificially to help extremely premature lambs develop to near maturity &#40;<a class="elsevierStyleCrossRef" href="#bb0205">Partridge et al&#46;&#44; 2017</a>&#41;&#46; Although this has not yet been achieved in humans&#44; in the future&#44; culture systems incorporated with a 3D printed vascularised umbilical cord and a functional artificial placenta <span class="elsevierStyleItalic">in vitro</span> could help extend <span class="elsevierStyleItalic">in vitro</span> embryo culture to later stages of development &#40;<a class="elsevierStyleCrossRef" href="#bb0365">Zhai et al&#46;&#44; 2021</a>&#41;&#46;</p><p id="p0300" class="elsevierStylePara elsevierViewall">Ultimately&#44; any decision to revise the 14-day rule must depend on whether the proposed changes can maintain the two main objectives of the rule&#58; to support research and to accommodate various moral concerns &#40;<a class="elsevierStyleCrossRef" href="#bb0150">Hyun et al&#46;&#44; 2016</a>&#41;&#46; It requires a careful assessment of the scientific feasibility and effective benefits of embryo research&#59; it needs extensive research on public opinion regarding embryos&#44; and a deliberative process that takes these elements into account&#46; It does not need positions that only consider the beneficence of the research and its technical feasibility&#46; This would be undemocratic and potentially a measure unsupported by a rigorous assessment of the science behind embryo research &#40;<a class="elsevierStyleCrossRef" href="#bb0050">Cavaliere&#44; 2017</a>&#41;&#46;</p></span></span></span><span id="s0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0135">Conclusions</span><p id="p0305" class="elsevierStylePara elsevierViewall">The extended culture of embryos up to day 14 of development is leading to a better understanding of the events that take place between implantation and gastrulation&#46; Despite this&#44; it is necessary to improve the current culture conditions in order to optimise the technique and reach the maximum number of days allowed by law&#44; as in most studies of this type&#44; the embryos are blocked before day 14&#46; Furthermore&#44; it should be borne in mind that access to and availability of human embryos is limited&#44; so other alternatives such as embryo development from stem cells should also be further explored&#46;</p><p id="p0310" class="elsevierStylePara elsevierViewall">Once the culture conditions up to day 14 have been optimised&#44; the idea of prolonging extended culture beyond day 14 is under debate&#44; since with the current knowledge and with the help of existing technology&#44; events such as gastrulation&#44; hitherto unknown in humans&#44; could be studied&#44; allowing a better understanding of human embryonic development at later stages&#46; In addition&#44; this could be applied to areas such as assisted reproduction when making decisions on the choice of the best embryos or on the knowledge of diseases that may affect pregnancy and lead to miscarriage&#44; for example&#46; All of this with clear moral and ethical boundaries&#46;</p></span><span id="s0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0140">Funding</span><p id="p0315" class="elsevierStylePara elsevierViewall">This manuscript has not been funded by any organization&#46;</p></span><span id="s0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0145">Conflict of interest</span><p id="p0320" class="elsevierStylePara elsevierViewall">The authors declare that there is no conflict of interest regarding the publication of this manuscript&#46;</p></span></span>"
    "textoCompletoSecciones" => array:1 [
      "secciones" => array:12 [
        0 => array:3 [
          "identificador" => "xres1786523"
          "titulo" => "Abstract"
          "secciones" => array:4 [
            0 => array:2 [
              "identificador" => "as0005"
              "titulo" => "Introduction"
            ]
            1 => array:2 [
              "identificador" => "as0010"
              "titulo" => "Materials and methods"
            ]
            2 => array:2 [
              "identificador" => "as0015"
              "titulo" => "Results and discussion"
            ]
            3 => array:2 [
              "identificador" => "as0020"
              "titulo" => "Conclusions"
            ]
          ]
        ]
        1 => array:2 [
          "identificador" => "xpalclavsec1566139"
          "titulo" => "Keywords"
        ]
        2 => array:2 [
          "identificador" => "xpalclavsec1566141"
          "titulo" => "Abbreviations"
        ]
        3 => array:3 [
          "identificador" => "xres1786524"
          "titulo" => "Resumen"
          "secciones" => array:4 [
            0 => array:2 [
              "identificador" => "as0025"
              "titulo" => "Introducci&#243;n"
            ]
            1 => array:2 [
              "identificador" => "as0030"
              "titulo" => "Materiales y m&#233;todos"
            ]
            2 => array:2 [
              "identificador" => "as0035"
              "titulo" => "Resultados y discusi&#243;n"
            ]
            3 => array:2 [
              "identificador" => "as0040"
              "titulo" => "Conclusiones"
            ]
          ]
        ]
        4 => array:2 [
          "identificador" => "xpalclavsec1566140"
          "titulo" => "Palabras clave"
        ]
        5 => array:2 [
          "identificador" => "s0005"
          "titulo" => "Introduction"
        ]
        6 => array:2 [
          "identificador" => "s0010"
          "titulo" => "Materials and methods"
        ]
        7 => array:3 [
          "identificador" => "s0015"
          "titulo" => "Results and discussion"
          "secciones" => array:2 [
            0 => array:2 [
              "identificador" => "s0020"
              "titulo" => "What is extended embryo culture up to 14 days and what are its conditions&#63;"
            ]
            1 => array:3 [
              "identificador" => "s0025"
              "titulo" => "The conditions and components of the extended culture medium are explained below"
              "secciones" => array:8 [
                0 => array:2 [
                  "identificador" => "s0030"
                  "titulo" => "Temperature"
                ]
                1 => array:2 [
                  "identificador" => "s0035"
                  "titulo" => "Gas environment"
                ]
                2 => array:2 [
                  "identificador" => "s0040"
                  "titulo" => "Culture media composition"
                ]
                3 => array:2 [
                  "identificador" => "s0045"
                  "titulo" => "What structures can we see with the culture up to day 14&#63;"
                ]
                4 => array:2 [
                  "identificador" => "s0050"
                  "titulo" => "Stem cells and culture"
                ]
                5 => array:2 [
                  "identificador" => "s0055"
                  "titulo" => "Potential applications in embryology"
                ]
                6 => array:2 [
                  "identificador" => "s0060"
                  "titulo" => "14-day limit"
                ]
                7 => array:2 [
                  "identificador" => "s0065"
                  "titulo" => "So&#44; should the 14-day limit be extended&#63;"
                ]
              ]
            ]
          ]
        ]
        8 => array:2 [
          "identificador" => "s0070"
          "titulo" => "Conclusions"
        ]
        9 => array:2 [
          "identificador" => "s0075"
          "titulo" => "Funding"
        ]
        10 => array:2 [
          "identificador" => "s0080"
          "titulo" => "Conflict of interest"
        ]
        11 => array:1 [
          "titulo" => "References"
        ]
      ]
    ]
    "pdfFichero" => "main.pdf"
    "tienePdf" => true
    "fechaRecibido" => "2022-02-23"
    "fechaAceptado" => "2022-05-19"
    "PalabrasClave" => array:2 [
      "en" => array:2 [
        0 => array:4 [
          "clase" => "keyword"
          "titulo" => "Keywords"
          "identificador" => "xpalclavsec1566139"
          "palabras" => array:5 [
            0 => "human embryo development"
            1 => "<span class="elsevierStyleItalic">in vitro</span> embryo development"
            2 => "pre-implantation"
            3 => "<span class="elsevierStyleItalic">in vitro</span> embryo culture"
            4 => "14-day-rule"
          ]
        ]
        1 => array:4 [
          "clase" => "abr"
          "titulo" => "Abbreviations"
          "identificador" => "xpalclavsec1566141"
          "palabras" => array:28 [
            0 => "2D"
            1 => "3D"
            2 => "ART"
            3 => "ATP"
            4 => "BSA"
            5 => "CO<span class="elsevierStyleInf">2</span>"
            6 => "DMEM&#47;F12"
            7 => "DNA"
            8 => "ESC"
            9 => "GATA6"
            10 => "HFE"
            11 => "HSA"
            12 => "ICM"
            13 => "ISSCR"
            14 => "ITS-X"
            15 => "IVC1"
            16 => "IVC2"
            17 => "IVF"
            18 => "Nanog"
            19 => "NCBI"
            20 => "NGS"
            21 => "O<span class="elsevierStyleInf">2</span>"
            22 => "OCT4"
            23 => "PKC"
            24 => "SBF"
            25 => "Sox17"
            26 => "TSC"
            27 => "XENC"
          ]
        ]
      ]
      "es" => array:1 [
        0 => array:4 [
          "clase" => "keyword"
          "titulo" => "Palabras clave"
          "identificador" => "xpalclavsec1566140"
          "palabras" => array:5 [
            0 => "desarrollo embrionario humano"
            1 => "desarrollo embrionario <span class="elsevierStyleItalic">in vitro</span>"
            2 => "pre-implantaci&#243;n"
            3 => "cultivo embrionario <span class="elsevierStyleItalic">in vitro</span>"
            4 => "regla de los 14 d&#237;as"
          ]
        ]
      ]
    ]
    "tieneResumen" => true
    "resumen" => array:2 [
      "en" => array:3 [
        "titulo" => "Abstract"
        "resumen" => "<span id="as0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0010">Introduction</span><p id="sp0025" class="elsevierStyleSimplePara elsevierViewall">The early stages of human embryonic development prior to implantation have been extensively studied to improve reproductive outcomes&#46; However&#44; subsequent events are unknown&#46; <span class="elsevierStyleItalic">In vitro</span> advances have allowed extended culture of embryos up to day 14&#44; considered the legal limit&#46; We aim to review aspects related culture conditions&#44; morphogenesis events&#44; potential applications of extended culture of human embryos as well as some ethical issues surrounding this topic&#46;</p></span> <span id="as0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0015">Materials and methods</span><p id="sp0030" class="elsevierStyleSimplePara elsevierViewall">This literature review was carried out using different databases and websites by screening articles&#46;</p></span> <span id="as0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0020">Results and discussion</span><p id="sp0035" class="elsevierStyleSimplePara elsevierViewall">Extended culture consists of culturing embryos up to day 14 of development&#46; This requires specific culture conditions of temperature and gas concentration&#44; as well as a medium composition that is different from that used in clinical practice in IVF laboratories&#46; In addition to having made it possible to document <span class="elsevierStyleItalic">in vitro</span> the morphological events and structures of the embryo after implantation&#44; the study of mosaic embryos at these hitherto hidden stages could provide valuable information for clinical practice&#46; All this has been achieved within the 14-day limit&#44; but due to scientific progress this can be considered restrictive and different reasons have been put forward why it should be extended&#46;</p></span> <span id="as0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0025">Conclusions</span><p id="sp0040" class="elsevierStyleSimplePara elsevierViewall">Although prolonged culture has provided a better understanding of the events occurring between implantation and gastrulation&#44; culture conditions must be improved to optimise the technique&#44; and once this is achieved&#44; consideration should be given to extending this limit&#46; Furthermore&#44; alternatives to the use of embryos&#44; such as the development of embryos from stem cells&#44; should be further explored&#46; All of this could have a future clinical application in terms of choosing the best embryos or finding out about diseases that affect pregnancy and lead to miscarriage&#46;</p></span>"
        "secciones" => array:4 [
          0 => array:2 [
            "identificador" => "as0005"
            "titulo" => "Introduction"
          ]
          1 => array:2 [
            "identificador" => "as0010"
            "titulo" => "Materials and methods"
          ]
          2 => array:2 [
            "identificador" => "as0015"
            "titulo" => "Results and discussion"
          ]
          3 => array:2 [
            "identificador" => "as0020"
            "titulo" => "Conclusions"
          ]
        ]
      ]
      "es" => array:3 [
        "titulo" => "Resumen"
        "resumen" => "<span id="as0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0035">Introducci&#243;n</span><p id="sp0045" class="elsevierStyleSimplePara elsevierViewall">Las primeras etapas del desarrollo embrionario humano antes de la implantaci&#243;n han sido ampliamente estudiadas para mejorar los resultados reproductivos&#46; Sin embargo&#44; se desconocen los eventos posteriores&#46; Los avances <span class="elsevierStyleItalic">in vitro</span> han permitido cultivar embriones hasta d&#237;a 14&#44; considerado el l&#237;mite legal&#46; Nuestro objetivo es hacer una revisi&#243;n sobre las condiciones de cultivo&#44; los eventos de morfog&#233;nesis&#44; las posibles aplicaciones del cultivo extendido de embriones humanos&#44; as&#237; como algunas cuestiones &#233;ticas que rodean este tema&#46;</p></span> <span id="as0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0040">Materiales y m&#233;todos</span><p id="sp0050" class="elsevierStyleSimplePara elsevierViewall">Esta revisi&#243;n bibliogr&#225;fica se ha realizado empleando diferentes bases de datos y p&#225;ginas web mediante el cribado de art&#237;culos&#46;</p></span> <span id="as0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0045">Resultados y discusi&#243;n</span><p id="sp0055" class="elsevierStyleSimplePara elsevierViewall">El cultivo extendido consiste en cultivar los embriones hasta d&#237;a 14&#46; Para ello se requieren unas condiciones de cultivo determinadas de temperatura y concentraci&#243;n de gases&#44; as&#237; como una composici&#243;n del medio que es diferente a la empleada en la pr&#225;ctica cl&#237;nica de los laboratorios de FIV&#46; Adem&#225;s de haber permitido documentar <span class="elsevierStyleItalic">in vitro</span> los eventos y estructuras morfol&#243;gicas del embri&#243;n tras su implantaci&#243;n&#44; el estudio de embriones mosaicos en estas etapas hasta ahora ocultas podr&#237;a dar informaci&#243;n valiosa para la pr&#225;ctica cl&#237;nica&#46; Todo esto se ha logrado dentro del l&#237;mite de 14 d&#237;as&#44; pero debido al avance cient&#237;fico puede considerarse restrictivo&#44; y se han expuesto diferentes motivos por los que deber&#237;a ampliarse&#46;</p></span> <span id="as0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="st0050">Conclusiones</span><p id="sp0060" class="elsevierStyleSimplePara elsevierViewall">Aunque el cultivo prolongado ha permitido comprender mejor los acontecimientos que ocurren entre la implantaci&#243;n y la gastrulaci&#243;n&#44; se deben mejorar las condiciones de cultivo para optimizar la t&#233;cnica&#44; y una vez conseguido&#44; plantear la ampliaci&#243;n de este l&#237;mite&#46; Adem&#225;s&#44; se deben seguir estudiando alternativas al uso de embriones como el desarrollo de embriones con c&#233;lulas madre&#46; Todo esto podr&#237;a tener una aplicaci&#243;n cl&#237;nica futura a la hora de elegir los mejores embriones o conocer enfermedades que afecten al embarazo y acaben en aborto&#46;</p></span>"
        "secciones" => array:4 [
          0 => array:2 [
            "identificador" => "as0025"
            "titulo" => "Introducci&#243;n"
          ]
          1 => array:2 [
            "identificador" => "as0030"
            "titulo" => "Materiales y m&#233;todos"
          ]
          2 => array:2 [
            "identificador" => "as0035"
            "titulo" => "Resultados y discusi&#243;n"
          ]
          3 => array:2 [
            "identificador" => "as0040"
            "titulo" => "Conclusiones"
          ]
        ]
      ]
    ]
    "multimedia" => array:4 [
      0 => array:8 [
        "identificador" => "f0005"
        "etiqueta" => "Figure 1"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr1.jpeg"
            "Alto" => 1154
            "Ancho" => 1778
            "Tamanyo" => 153277
          ]
        ]
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "al0005"
            "detalle" => "Figure "
            "rol" => "short"
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="sp0005" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleBold">Extended culture procedure up to day 14&#46;</span> The embryos are first thawed and then placed in culture medium for two hours for reexpansion&#46; The next step is to remove the zona pellucida with tyrodes acid and then transfer them to the culture plate for culture until day 14 of development&#46; Each day they can be observed under the microscope and their evolution can be evaluated&#46;</p>"
        ]
      ]
      1 => array:8 [
        "identificador" => "f0010"
        "etiqueta" => "Figure 2"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr2.jpeg"
            "Alto" => 674
            "Ancho" => 1778
            "Tamanyo" => 119460
          ]
        ]
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "al0010"
            "detalle" => "Figure "
            "rol" => "short"
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="sp0010" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleBold">Events of human implantation morphogenesis&#46;</span> The embryo formed by the ICM &#40;inner cell mass&#41; and the trophectoderm is shown first&#46; On the one hand&#44; from the ICM&#44; the epiblast and hypoblast segregate&#44; forming the bilaminar disc&#46; In the epiblast&#44; a cavity called the pro-amniotic cavity begins to appear&#44; which will later give rise to the amniotic cavity&#44; and the hypoblast extends to form the yolk sac&#46; On the other hand&#44; the trophectoderm differentiates into cytotrophoblast and syncytiotrophoblast&#46;</p>"
        ]
      ]
      2 => array:8 [
        "identificador" => "f0015"
        "etiqueta" => "Figure 3"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr3.jpeg"
            "Alto" => 1012
            "Ancho" => 1611
            "Tamanyo" => 117760
          ]
        ]
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "al0015"
            "detalle" => "Figure "
            "rol" => "short"
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="sp0015" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleBold">Non-integrated and integrated Stem cell-based embryo models&#46;</span> The upper figure of the schematic corresponds to an amniotic sac-like structure obtained from ESC&#46; The second represents the elongated gastruloid with an anterior-posterior axis&#44; also obtained from ESC&#46; The third structure represents the embryoid&#44; which has been obtained from ESC&#44; XENC and TSC&#46; And the last structure is known as the blastoid&#44; made up of cells from ESC and TSC&#46; &#40;ESC&#58; embryonic stem cells&#59; XENC&#58; extraembryonic endoderm cells&#59; TSC&#58; trophectoderm stem cells&#41; &#40;adapted from <a class="elsevierStyleCrossRef" href="#bb0245">Rossant and Tam&#44; 2021</a>&#41;&#46;</p>"
        ]
      ]
      3 => array:8 [
        "identificador" => "t0005"
        "etiqueta" => "Table 1"
        "tipo" => "MULTIMEDIATABLA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "al0020"
            "detalle" => "Table "
            "rol" => "short"
          ]
        ]
        "tabla" => array:1 [
          "tablatextoimagen" => array:1 [
            0 => array:1 [
              "tabla" => array:1 [
                0 => """
                  <table border="0" frame="\n
                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Advanced DMEM&#47;F12&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Concentration &#40;mg&#47;L&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Concentration &#40;mg&#47;L&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Non-essential amino acids&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Essential amino acids&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Glycine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">18&#46;75&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Arginine hydrochloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">147&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Alanine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">4&#46;45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Histidine hydrochloride-H<span class="elsevierStyleInf">2</span>O&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">31&#46;48&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Asparagine-H<span class="elsevierStyleInf">2</span>O&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">7&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Isoleucine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">54&#46;47&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Aspartic acid&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">6&#46;65&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Leucine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">59&#46;05&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Cysteine hydrochloride-H<span class="elsevierStyleInf">2</span>O&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">17&#46;56&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Lysine hydrochloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">91&#46;25&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Cysteine 2HCl&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">31&#46;29&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Methionine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">17&#46;24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Glutamic acid&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">7&#46;35&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Phenylalanine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">35&#46;48&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Proline&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">17&#46;25&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Threonine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">53&#46;45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Serine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">26&#46;25&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Tryptophan&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">9&#46;02&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Tyrosine disodium salt dihydrate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">55&#46;79&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">L-Valine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">52&#46;85&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Inorganic salts&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Vitamins&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Calcium chloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">116&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Ascorbic acid phosphate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Cupric sulphate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;0013&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Biotin&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;0035&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Ferric nitrate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;05&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Choline chloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">8&#46;98&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Ferric sulphate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;417&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">D-Calcium pantothenate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Magnesium chloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">28&#46;64&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Folic acid&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;65&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Magnesium sulphate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">48&#46;84&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Niacinamide&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;02&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Potassium chloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">311&#46;8&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Pyridoxine hydrochloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Sodium bicarbonate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2438&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Riboflavin&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;219&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Sodium chloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">6995&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Thiamine hydrochloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;17&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Sodium phosphate dibasic&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">71&#46;02&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Vitamin B12&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;68&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Sodium phosphate monobasic&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">62&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">i-Inositol&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">132&#46;6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Zinc sulphate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;864&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Other components&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Ammonium metavanadate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">3&#46;0E-4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">D-Glucose &#40;Dextrose&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">3151&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Manganous chloride&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">5&#46;0E-5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Ethanolamine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1&#46;9&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Sodium selenite&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;005&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Hypoxanthine Na&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;39&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Proteins&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Linoleic acid&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;042&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">AlbuMAX&#174; II&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">400&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Lipoic acid&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;105&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Human Transferrin&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">7&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Phenol red&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">8&#46;1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Insulin Recombinant Full Chain&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">10&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Putrescine 2HCl&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;081&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Reducing agents&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Sodium pyruvate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">110&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Glutathione&#44; monosodium&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Thymidine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;365&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleBold">ITS-X</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Proteins&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Insulin&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1000&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Transferrin&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">550&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Trace elements&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Sodium selenite&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;67&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">Other components&#58;</span></span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Ethanolamine&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">200&#46;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
              ]
            ]
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="sp0020" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleBold">Composition and concentrations of basal medium Advanced DMEM&#47;F12 and ITS-X&#46;</span> This table shows the different components that make up the basal medium Advanced DMEM&#47;F12&#44; as well as ITS-X&#44; all with their respective concentrations in mg&#47;L&#46; Data obtained from <a class="elsevierStyleCrossRefs" href="#bb0285">ThermoFisher Scientific &#40;2014a&#44; 2014b&#41;</a>&#46;</p>"
        ]
      ]
    ]
    "bibliografia" => array:2 [
      "titulo" => "References"
      "seccion" => array:1 [
        0 => array:2 [
          "identificador" => "bs0005"
          "bibliografiaReferencia" => array:74 [
            0 => array:3 [
              "identificador" => "bb0005"
              "etiqueta" => "Aach et al&#46;&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Addressing the ethical issues raised by synthetic human entities with embryo-like features"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [
                            0 => "J&#46; Aach"
                            1 => "J&#46; Lunshof"
                            2 => "E&#46; Iyer"
                            3 => "G&#46;M&#46; Church"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:3 [
                        "tituloSerie" => "eLife Sci&#46;"
                        "fecha" => "2017"
                        "volumen" => "6"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            1 => array:3 [
              "identificador" => "bb0010"
              "etiqueta" => "Appleby and Bredenoord&#44; 2018"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Should the 14-day rule for embryo research become the 28-day rule&#63;"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "J&#46;B&#46; Appleby"
                            1 => "A&#46;L&#46; Bredenoord"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:4 [
                        "tituloSerie" => "EMBO Mol&#46; Med&#46;"
                        "fecha" => "2018"
                        "volumen" => "10"
                        "numero" => "9"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            2 => array:3 [
              "identificador" => "bb0015"
              "etiqueta" => "Baak et al&#46;&#44; 2019"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Temperature of embryo culture for assisted reproduction"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [
                            0 => "N&#46;A&#46; Baak"
                            1 => "A&#46;E&#46;P&#46; Cantineau"
                            2 => "C&#46; Farquhar"
                            3 => "D&#46;R&#46; Brison"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:5 [
                        "tituloSerie" => "Cochrane Database Syst&#46; Rev&#46;"
                        "fecha" => "2019"
                        "volumen" => "9"
                        "paginaInicial" => "1"
                        "paginaFinal" => "34"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            3 => array:3 [
              "identificador" => "bb0020"
              "etiqueta" => "Beccari et al&#46;&#44; 2018"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Multi-axialself-organization properties of mouse embryonic stem cells into gastruloids"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:9 [
                            0 => "L&#46; Beccari"
                            1 => "N&#46; Moris"
                            2 => "M&#46; Girgin"
                            3 => "D&#46;A&#46; Turner"
                            4 => "P&#46; Baillie-Johnson"
                            5 => "A&#46;C&#46; Cossy"
                            6 => "M&#46;P&#46; Lutolf"
                            7 => "D&#46; Duboule"
                            8 => "A&#46;M&#46; Arias"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46;"
                        "fecha" => "2018"
                        "volumen" => "562"
                        "numero" => "7726"
                        "paginaInicial" => "272"
                        "paginaFinal" => "276"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            4 => array:3 [
              "identificador" => "bb0025"
              "etiqueta" => "Bedzhov et al&#46;&#44; 2014"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "<span class="elsevierStyleItalic">In vitro</span> culture of mouse blastocysts beyond the implantation stages"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [
                            0 => "I&#46; Bedzhov"
                            1 => "C&#46;Y&#46; Leung"
                            2 => "M&#46; Bialecka"
                            3 => "M&#46; Zernicka-Goetz"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/nprot.2014.186"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Protoc&#46;"
                        "fecha" => "2014"
                        "volumen" => "9"
                        "numero" => "12"
                        "paginaInicial" => "2732"
                        "paginaFinal" => "2739"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/25356584"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            5 => array:3 [
              "identificador" => "bb0030"
              "etiqueta" => "Blackshaw and Rodger&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Why we should not extend the 14-day rule"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "B&#46;P&#46; Blackshaw"
                            1 => "D&#46; Rodger"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1136/medethics-2021-107317"
                      "Revista" => array:7 [
                        "tituloSerie" => "J&#46; Med&#46; Ethics"
                        "fecha" => "2021"
                        "volumen" => "47"
                        "numero" => "10"
                        "paginaInicial" => "712"
                        "paginaFinal" => "714"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/34112713"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            6 => array:3 [
              "identificador" => "bb0035"
              "etiqueta" => "Bolton et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Mouse model of chromosome mosaicism reveals lineage-specific depletion of aneuploid cells and normal developmental potential"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:8 [
                            0 => "H&#46; Bolton"
                            1 => "S&#46;J&#46; Graham"
                            2 => "N&#46; Van der Aa"
                            3 => "P&#46; Kumar"
                            4 => "K&#46; Theunis"
                            5 => "E&#46; Fernandez Gallardo"
                            6 => "T&#46; Voet"
                            7 => "M&#46; Zernicka-Goetz"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46; Commun&#46;"
                        "fecha" => "2016"
                        "volumen" => "7"
                        "numero" => "1"
                        "paginaInicial" => "1"
                        "paginaFinal" => "12"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            7 => array:3 [
              "identificador" => "bb0040"
              "etiqueta" => "Bredenoord et al&#46;&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Human tissues in a dish&#58; the research and ethical implications of organoid technology"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "A&#46;L&#46; Bredenoord"
                            1 => "H&#46; Clevers"
                            2 => "J&#46;A&#46; Knoblich"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1126/science.aaf9414"
                      "Revista" => array:6 [
                        "tituloSerie" => "Science&#46;"
                        "fecha" => "2017"
                        "volumen" => "355"
                        "numero" => "6322"
                        "paginaInicial" => "eaaf9414"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28104841"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            8 => array:3 [
              "identificador" => "bb0045"
              "etiqueta" => "Castelyn&#44; 2020"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Embryo experimentation&#58; is there a case for moving beyond the &#8216;14-day rule&#8217;"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "G&#46; Castelyn"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Monash Bioeth&#46; Rev&#46;"
                        "fecha" => "2020"
                        "volumen" => "38"
                        "numero" => "2"
                        "paginaInicial" => "181"
                        "paginaFinal" => "196"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            9 => array:3 [
              "identificador" => "bb0050"
              "etiqueta" => "Cavaliere&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "A 14-day limit for bioethics&#58; the debate over human embryo research"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "G&#46; Cavaliere"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1186/s12910-016-0160-y"
                      "Revista" => array:7 [
                        "tituloSerie" => "BMC Med&#46; Ethics&#46;"
                        "fecha" => "2017"
                        "volumen" => "18"
                        "numero" => "1"
                        "paginaInicial" => "1"
                        "paginaFinal" => "12"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28077127"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            10 => array:3 [
              "identificador" => "bb0055"
              "etiqueta" => "Chattopadhyay et al&#46;&#44; 2003"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Polyamines protect <span class="elsevierStyleItalic">Escherichia coli</span> cells from the toxic effect of oxygen"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "M&#46;K&#46; Chattopadhyay"
                            1 => "C&#46;W&#46; Tabor"
                            2 => "H&#46; Tabor"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1073/pnas.2627990100"
                      "Revista" => array:7 [
                        "tituloSerie" => "Proc&#46; Natl&#46; Acad&#46; Sci&#46; U&#46; S&#46; A&#46;"
                        "fecha" => "2003"
                        "volumen" => "100"
                        "numero" => "5"
                        "paginaInicial" => "2261"
                        "paginaFinal" => "2265"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/12591940"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            11 => array:3 [
              "identificador" => "bb0060"
              "etiqueta" => "Christianson et al&#46;&#44; 2014"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Embryo catheter loading and embryo culture techniques&#58; results of a worldwide web-based survey"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:8 [
                            0 => "M&#46;S&#46; Christianson"
                            1 => "Y&#46; Zhao"
                            2 => "G&#46; Shoham"
                            3 => "I&#46; Granot"
                            4 => "A&#46; Safran"
                            5 => "A&#46; Khafagy"
                            6 => "M&#46; Leong"
                            7 => "Z&#46; Shoham"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1007/s10815-014-0250-z"
                      "Revista" => array:7 [
                        "tituloSerie" => "J&#46; Assist&#46; Reprod&#46; Genet&#46;"
                        "fecha" => "2014"
                        "volumen" => "31"
                        "numero" => "8"
                        "paginaInicial" => "1029"
                        "paginaFinal" => "1036"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/24913025"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            12 => array:3 [
              "identificador" => "bb0065"
              "etiqueta" => "De Los Santos et al&#46;&#44; 2013"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Reduced oxygen tension improves embryo quality but not clinical pregnancy rates&#58; a randomized clinical study into ovum donation cycles"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:8 [
                            0 => "M&#46;J&#46; De Los Santos"
                            1 => "P&#46; G&#225;miz"
                            2 => "C&#46; Albert"
                            3 => "A&#46; Gal&#225;n"
                            4 => "T&#46; Viloria"
                            5 => "S&#46; P&#233;rez"
                            6 => "J&#46;L&#46; Romero"
                            7 => "J&#46; Remoh&#237;"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.fertnstert.2013.03.044"
                      "Revista" => array:7 [
                        "tituloSerie" => "Fertil&#46; Steril&#46;"
                        "fecha" => "2013"
                        "volumen" => "100"
                        "numero" => "2"
                        "paginaInicial" => "402"
                        "paginaFinal" => "407"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23628109"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            13 => array:3 [
              "identificador" => "bb0070"
              "etiqueta" => "Deglincerti et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Self-organization of the <span class="elsevierStyleItalic">in vitro</span> attached human embryo"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:6 [
                            0 => "A&#46; Deglincerti"
                            1 => "G&#46;F&#46; Croft"
                            2 => "L&#46;N&#46; Pietila"
                            3 => "M&#46; Zernicka-Goetz"
                            4 => "E&#46;D&#46; Siggia"
                            5 => "A&#46;H&#46; Brivanlou"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46;"
                        "fecha" => "2016"
                        "volumen" => "533"
                        "numero" => "7602"
                        "paginaInicial" => "251"
                        "paginaFinal" => "254"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            14 => array:3 [
              "identificador" => "bb0075"
              "etiqueta" => "Edwards et al&#46;&#44; 1969"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Early stages of fertilization <span class="elsevierStyleItalic">in vitro</span> of human oocytes matured <span class="elsevierStyleItalic">in vitro</span>"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "R&#46;G&#46; Edwards"
                            1 => "B&#46;D&#46; Bavister"
                            2 => "P&#46;C&#46; Steptoe"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46;"
                        "fecha" => "1969"
                        "volumen" => "221"
                        "numero" => "5181"
                        "paginaInicial" => "632"
                        "paginaFinal" => "635"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            15 => array:3 [
              "identificador" => "bb0080"
              "etiqueta" => "Edwards et al&#46;&#44; 1970"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Fertilization and cleavage <span class="elsevierStyleItalic">in vitro</span> of preovulator human oocytes"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "R&#46;G&#46; Edwards"
                            1 => "P&#46;C&#46; Steptoe"
                            2 => "J&#46;M&#46; Purdy"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46;"
                        "fecha" => "1970"
                        "volumen" => "227"
                        "numero" => "5265"
                        "paginaInicial" => "1307"
                        "paginaFinal" => "1309"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            16 => array:3 [
              "identificador" => "bb0085"
              "etiqueta" => "Enders et al&#46;&#44; 1986"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Differentiation of the embryonic disc&#44; amnion&#44; and yolk sac in the rhesus monkey"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "A&#46;C&#46; Enders"
                            1 => "S&#46; Schlafke"
                            2 => "A&#46;G&#46; Hendrickx"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1002/aja.1001770205"
                      "Revista" => array:7 [
                        "tituloSerie" => "Am&#46; J&#46; Anat&#46;"
                        "fecha" => "1986"
                        "volumen" => "177"
                        "numero" => "2"
                        "paginaInicial" => "161"
                        "paginaFinal" => "185"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/3788819"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            17 => array:3 [
              "identificador" => "bb0090"
              "etiqueta" => "Etoc et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "A balance between secreted inhibitors and Edge sensing controls gastruloid self-organization"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:8 [
                            0 => "F&#46; Etoc"
                            1 => "J&#46; Metzger"
                            2 => "A&#46; Ruzo"
                            3 => "C&#46; Kirst"
                            4 => "A&#46; Yoney"
                            5 => "M&#46;Z&#46; Ozair"
                            6 => "A&#46;H&#46; Brivanlou"
                            7 => "E&#46;D&#46; Siggia"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.devcel.2016.09.016"
                      "Revista" => array:7 [
                        "tituloSerie" => "Dev&#46; Cell"
                        "fecha" => "2016"
                        "volumen" => "39"
                        "numero" => "3"
                        "paginaInicial" => "302"
                        "paginaFinal" => "315"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27746044"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            18 => array:3 [
              "identificador" => "bb0095"
              "etiqueta" => "F&#252;rstova et al&#46;&#44; 2008"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Comparison of the effect of individual saturated and unsaturated fatty acids on cell growth and death induction in the human pancreatic &#946;-cell line NES2Y"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [
                            0 => "V&#46; F&#252;rstova"
                            1 => "T&#46; Kopska"
                            2 => "R&#46;F&#46; James"
                            3 => "J&#46; Kovar"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.lfs.2007.12.023"
                      "Revista" => array:7 [
                        "tituloSerie" => "Life Sci&#46;"
                        "fecha" => "2008"
                        "volumen" => "82"
                        "numero" => "13-14"
                        "paginaInicial" => "684"
                        "paginaFinal" => "691"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/18272185"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            19 => array:3 [
              "identificador" => "bb0100"
              "etiqueta" => "Greco et al&#46;&#44; 2015"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Healthy babies after intrauterine transfer of mosaic aneuploid blastocysts"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "E&#46; Greco"
                            1 => "M&#46; Minasi"
                            2 => "F&#46; Fiorentino"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1056/NEJMc1500421"
                      "Revista" => array:7 [
                        "tituloSerie" => "N&#46; Engl&#46; J&#46; Med&#46;"
                        "fecha" => "2015"
                        "volumen" => "373"
                        "numero" => "21"
                        "paginaInicial" => "2089"
                        "paginaFinal" => "2090"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/26581010"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            20 => array:3 [
              "identificador" => "bb0105"
              "etiqueta" => "Green et al&#46;&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Don&#8217;t abandon 14-day limit on embryo research&#44; it makes sense"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "R&#46;M&#46; Green"
                            1 => "M&#46;D&#46; West"
                            2 => "L&#46; Hayflick"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/d41586-021-01595-7"
                      "Revista" => array:6 [
                        "tituloSerie" => "Nature&#46;"
                        "fecha" => "2021"
                        "volumen" => "594"
                        "numero" => "7863"
                        "paginaInicial" => "333"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/34131341"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            21 => array:3 [
              "identificador" => "bb0110"
              "etiqueta" => "Halloran et al&#46;&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Arginine&#44; agmatine&#44; and polyamines&#58; key regulators of conceptus development in mammals"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [
                            0 => "K&#46;M&#46; Halloran"
                            1 => "C&#46; Stenhouse"
                            2 => "G&#46; Wu"
                            3 => "F&#46;W&#46; Bazer"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "LibroEditado" => array:6 [
                        "editores" => "G&#46;Wu"
                        "titulo" => "Amino Acids in Nutrition and Health"
                        "paginaInicial" => "85"
                        "paginaFinal" => "105"
                        "serieTitulo" => "Advances in Experimental Medicine and Biology"
                        "serieFecha" => "2021"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            22 => array:3 [
              "identificador" => "bb0115"
              "etiqueta" => "Harrison et al&#46;&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Assembly of embryonic and extra-embryonic stem cells to mimic embryogeneis <span class="elsevierStyleItalic">in vitro</span>"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:5 [
                            0 => "S&#46;E&#46; Harrison"
                            1 => "B&#46; Sozen"
                            2 => "N&#46; Christodoulou"
                            3 => "C&#46; Kyprianou"
                            4 => "M&#46; Zernicka-Goetz"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1126/science.aal1810"
                      "Revista" => array:5 [
                        "tituloSerie" => "Sci&#46;"
                        "fecha" => "2017"
                        "volumen" => "356"
                        "numero" => "6334"
                        "paginaInicial" => "eaal1810"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            23 => array:3 [
              "identificador" => "bb0120"
              "etiqueta" => "Hertig et al&#46;&#44; 1956"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "A description of 34 human ova within the first 17 days of development"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "A&#46;T&#46; Hertig"
                            1 => "J&#46; Rock"
                            2 => "E&#46;C&#46; Adams"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1002/aja.1000980306"
                      "Revista" => array:7 [
                        "tituloSerie" => "Am&#46; J&#46; Anat&#46;"
                        "fecha" => "1956"
                        "volumen" => "98"
                        "numero" => "3"
                        "paginaInicial" => "435"
                        "paginaFinal" => "493"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/13362122"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            24 => array:3 [
              "identificador" => "bb0125"
              "etiqueta" => "Hong et al&#46;&#44; 2014"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Examining the temperature of embryo culture in <span class="elsevierStyleItalic">in vitro</span> fertilization&#58; a randomized controlled trial comparing traditional core temperature &#40;37&#176;C&#41; to a more physiologic&#44; cooler temperature &#40;36&#176;C&#41;"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:5 [
                            0 => "K&#46;H&#46; Hong"
                            1 => "H&#46; Lee"
                            2 => "E&#46;J&#46; Forman"
                            3 => "K&#46;M&#46; Upham"
                            4 => "R&#46;T&#46; Scott Jr&#46;"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.fertnstert.2014.06.009"
                      "Revista" => array:7 [
                        "tituloSerie" => "Fertil&#46; Steril&#46;"
                        "fecha" => "2014"
                        "volumen" => "102"
                        "numero" => "3"
                        "paginaInicial" => "767"
                        "paginaFinal" => "773"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/25044079"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            25 => array:3 [
              "identificador" => "bb0130"
              "etiqueta" => "Human Fertilization and Embryology Act&#44; 1990"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "UK Legislation&#44; passed 1st November 1990&#46; Her Majesty&#8217;s Stationery Office &#40;HMSO&#41;"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "Human Fertilization and Embryology Act"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Libro" => array:1 [
                        "fecha" => "1990"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            26 => array:3 [
              "identificador" => "bb0135"
              "etiqueta" => "Human Fertilization and Embryology Act&#44; 2008"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "UK Legislation&#44; passed 13th November 2008"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "Human Fertilization and Embryology Act"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "LibroEditado" => array:2 [
                        "titulo" => "Her Majesty&#8217;s Stationery Office &#40;HMSO&#41;"
                        "serieFecha" => "2008"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            27 => array:3 [
              "identificador" => "bb0140"
              "etiqueta" => "Hurlbut et al&#46;&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Revisiting the Warnock rule"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:15 [
                            0 => "J&#46;B&#46; Hurlbut"
                            1 => "I&#46; Hyun"
                            2 => "A&#46;D&#46; Levine"
                            3 => "R&#46; Lovell-Badge"
                            4 => "J&#46;E&#46; Lunshof"
                            5 => "K&#46;R&#46; Matthews"
                            6 => "P&#46; Mills"
                            7 => "A&#46; Murdoch"
                            8 => "M&#46;F&#46; Pera"
                            9 => "C&#46;T&#46; Scott"
                            10 => "J&#46; Tizzard"
                            11 => "M&#46; Warnock"
                            12 => "M&#46; Zernicka-Goetz"
                            13 => "Q&#46; Zhou"
                            14 => "L&#46; Zoloth"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/nbt.4015"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Biotechnol&#46;"
                        "fecha" => "2017"
                        "volumen" => "35"
                        "numero" => "11"
                        "paginaInicial" => "1029"
                        "paginaFinal" => "1042"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/29121021"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            28 => array:3 [
              "identificador" => "bb0145"
              "etiqueta" => "Hussain et al&#46;&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Exploring polyamines&#58; functions in embryo&#47;fetal development"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:5 [
                            0 => "T&#46; Hussain"
                            1 => "W&#46; Ren"
                            2 => "N&#46; Rahu"
                            3 => "D&#46;H&#46; Kalhoro"
                            4 => "Y&#46; Yin"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.aninu.2016.12.002"
                      "Revista" => array:7 [
                        "tituloSerie" => "Anim&#46; Nutr&#46;"
                        "fecha" => "2017"
                        "volumen" => "3"
                        "numero" => "1"
                        "paginaInicial" => "7"
                        "paginaFinal" => "10"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/29767087"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            29 => array:3 [
              "identificador" => "bb0150"
              "etiqueta" => "Hyun et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Embryology policy&#58; revisit the 14-day rule"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "I&#46; Hyun"
                            1 => "A&#46; Wilkerson"
                            2 => "J&#46; Johnston"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46; N&#46;"
                        "fecha" => "2016"
                        "volumen" => "533"
                        "numero" => "7602"
                        "paginaInicial" => "169"
                        "paginaFinal" => "171"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            30 => array:3 [
              "identificador" => "bb0155"
              "etiqueta" => "Hyun et al&#46;&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Human embryo research beyond the primitive streak"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:5 [
                            0 => "I&#46; Hyun"
                            1 => "A&#46;L&#46; Bredenoord"
                            2 => "J&#46; Briscoe"
                            3 => "S&#46; Klipstein"
                            4 => "T&#46; Tan"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1126/science.abf3751"
                      "Revista" => array:7 [
                        "tituloSerie" => "Science&#46;"
                        "fecha" => "2021"
                        "volumen" => "371"
                        "numero" => "6533"
                        "paginaInicial" => "998"
                        "paginaFinal" => "1000"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/33674483"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            31 => array:3 [
              "identificador" => "bb0160"
              "etiqueta" => "Igarashi and Kashiwagi&#44; 2000"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Polyamines&#58; mysterious modulators of cellular functions"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "K&#46; Igarashi"
                            1 => "K&#46; Kashiwagi"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1006/bbrc.2000.2601"
                      "Revista" => array:7 [
                        "tituloSerie" => "Biochem&#46; Biophys&#46; Res&#46; Commun&#46;"
                        "fecha" => "2000"
                        "volumen" => "271"
                        "numero" => "3"
                        "paginaInicial" => "559"
                        "paginaFinal" => "564"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/10814501"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            32 => array:3 [
              "identificador" => "bb0165"
              "etiqueta" => "Kirkegaard et al&#46;&#44; 2013"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "K&#46; Kirkegaard"
                            1 => "J&#46;J&#46; Hindkjaer"
                            2 => "H&#46;J&#46; Ingerslev"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.fertnstert.2012.11.028"
                      "Revista" => array:7 [
                        "tituloSerie" => "Fertil&#46; Steril&#46;"
                        "fecha" => "2013"
                        "volumen" => "99"
                        "numero" => "3"
                        "paginaInicial" => "738"
                        "paginaFinal" => "744"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23245683"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            33 => array:3 [
              "identificador" => "bb0170"
              "etiqueta" => "Lusine&#44; 2020"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "How good are we at modeling implantation&#63;"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "A&#46; Lusine"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1007/s10815-019-01667-8"
                      "Revista" => array:7 [
                        "tituloSerie" => "J&#46; Assist&#46; Reprod&#46; Genet&#46;"
                        "fecha" => "2020"
                        "volumen" => "37"
                        "numero" => "1"
                        "paginaInicial" => "17"
                        "paginaFinal" => "19"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/31863287"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            34 => array:3 [
              "identificador" => "bb0175"
              "etiqueta" => "McCully&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "The time has come to extend the 14-day limit"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "S&#46; McCully"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:5 [
                        "tituloSerie" => "J&#46; Med&#46; Ethics"
                        "fecha" => "2021"
                        "volumen" => "47"
                        "numero" => "12"
                        "paginaInicial" => "e66"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            35 => array:3 [
              "identificador" => "bb0180"
              "etiqueta" => "McLaren&#44; 1984"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Where to draw the line"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "A&#46; McLaren"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:5 [
                        "tituloSerie" => "P&#46; Roy&#46; Inst&#46;"
                        "fecha" => "1984"
                        "volumen" => "56"
                        "paginaInicial" => "101"
                        "paginaFinal" => "121"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            36 => array:3 [
              "identificador" => "bb0185"
              "etiqueta" => "M&#233;n&#233;zo et al&#46;&#44; 2013"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "New insights into human pre-implantation metabolism <span class="elsevierStyleItalic">in vivo</span> and <span class="elsevierStyleItalic">in vitro</span>"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "Y&#46; M&#233;n&#233;zo"
                            1 => "I&#46; Lichtblau"
                            2 => "K&#46; Elder"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1007/s10815-013-9953-9"
                      "Revista" => array:7 [
                        "tituloSerie" => "J&#46; Assist&#46; Reprod&#46; Genet&#46;"
                        "fecha" => "2013"
                        "volumen" => "30"
                        "numero" => "3"
                        "paginaInicial" => "293"
                        "paginaFinal" => "303"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23430228"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            37 => array:3 [
              "identificador" => "bb0190"
              "etiqueta" => "Nastri et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Low versus atmospheric oxygen tension for embryo culture in assisted reproduction&#58; a systematic review and meta-analysis"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:9 [
                            0 => "C&#46;O&#46; Nastri"
                            1 => "B&#46;N&#46; N&#243;brega"
                            2 => "D&#46;M&#46; Teixeira"
                            3 => "J&#46; Amorim"
                            4 => "L&#46;M&#46;M&#46; Diniz"
                            5 => "M&#46;W&#46;P&#46; Barbosa"
                            6 => "V&#46;S&#46;I&#46; Giorgi"
                            7 => "V&#46;N&#46; Pileggi"
                            8 => "W&#46;P&#46; Martins"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.fertnstert.2016.02.037"
                      "Revista" => array:7 [
                        "tituloSerie" => "Fertil&#46; Steril&#46;"
                        "fecha" => "2016"
                        "volumen" => "106"
                        "numero" => "1"
                        "paginaInicial" => "95"
                        "paginaFinal" => "104 e117"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27012651"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            38 => array:3 [
              "identificador" => "bb0195"
              "etiqueta" => "Niu et al&#46;&#44; 2019"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Dissecting primate early post-implantation development using long-term <span class="elsevierStyleItalic">in vitro</span> embryo culture"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:23 [
                            0 => "Y&#46; Niu"
                            1 => "N&#46; Sun"
                            2 => "Y&#46; Li"
                            3 => "Z&#46; Huang"
                            4 => "J&#46; Wu"
                            5 => "C&#46; Si"
                            6 => "X&#46; Dai"
                            7 => "C&#46; Liu"
                            8 => "J&#46; Wei"
                            9 => "L&#46; Liu"
                            10 => "S&#46; Feng"
                            11 => "Y&#46; Kang"
                            12 => "W&#46; Si"
                            13 => "H&#46; Wang"
                            14 => "E&#46; Zhang"
                            15 => "L&#46; Zhao"
                            16 => "Z&#46; Li"
                            17 => "X&#46; Luo"
                            18 => "G&#46; Cui"
                            19 => "G&#46; Peng"
                            20 => "J&#46;C&#46; Izpis&#250;a Belmonte"
                            21 => "W&#46; Ji"
                            22 => "T&#46; Tan"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1126/science.aaw5754"
                      "Revista" => array:4 [
                        "tituloSerie" => "Sci&#46;"
                        "fecha" => "2019"
                        "volumen" => "366"
                        "numero" => "6467"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            39 => array:3 [
              "identificador" => "bb0200"
              "etiqueta" => "Nuffield Council on Bioethics&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Human embryo culture"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "Nuffield Council on Bioethics"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Libro" => array:3 [
                        "fecha" => "2017"
                        "editorial" => "Nuffield Council on Bioethics"
                        "editorialLocalizacion" => "London&#44; UK"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            40 => array:3 [
              "identificador" => "bb0205"
              "etiqueta" => "Partridge et al&#46;&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "An extra-uterine system to physiologically support the extreme premature lamb"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:18 [
                            0 => "E&#46;A&#46; Partridge"
                            1 => "M&#46;G&#46; Davey"
                            2 => "M&#46;A&#46; Hornick"
                            3 => "P&#46;E&#46; McGovern"
                            4 => "A&#46;Y&#46; Mejaddam"
                            5 => "J&#46;D&#46; Vrecenak"
                            6 => "C&#46; Mesas-Burgos"
                            7 => "A&#46; Olive"
                            8 => "R&#46;C&#46; Caskey"
                            9 => "T&#46;R&#46; Weiland"
                            10 => "J&#46; Han"
                            11 => "A&#46;J&#46; Schupper"
                            12 => "J&#46;T&#46; Connelly"
                            13 => "K&#46;C&#46; Dysart"
                            14 => "J&#46; Rychik"
                            15 => "H&#46;L&#46; Hedrick"
                            16 => "W&#46;H&#46; Peranteau"
                            17 => "A&#46;W&#46; Flake"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/s41467-016-0009-6"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Commun&#46;"
                        "fecha" => "2017"
                        "volumen" => "8"
                        "numero" => "1"
                        "paginaInicial" => "1"
                        "paginaFinal" => "16"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28232747"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            41 => array:3 [
              "identificador" => "bb0210"
              "etiqueta" => "Pawar and Jump&#44; 2003"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Unsaturated fatty acid regulation of peroxisome proliferator-activated receptor &#945; activity in rat primary hepatocytes"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "A&#46; Pawar"
                            1 => "D&#46;B&#46; Jump"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1074/jbc.M306238200"
                      "Revista" => array:7 [
                        "tituloSerie" => "J&#46; Biol&#46; Chem&#46;"
                        "fecha" => "2003"
                        "volumen" => "278"
                        "numero" => "38"
                        "paginaInicial" => "35931"
                        "paginaFinal" => "35939"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/12853447"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            42 => array:3 [
              "identificador" => "bb0215"
              "etiqueta" => "Pera&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Human embryo research and the 14-day rule"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "M&#46;F&#46; Pera"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1242/dev.151191"
                      "Revista" => array:7 [
                        "tituloSerie" => "Development&#46;"
                        "fecha" => "2017"
                        "volumen" => "144"
                        "numero" => "11"
                        "paginaInicial" => "1923"
                        "paginaFinal" => "1925"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/28559237"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            43 => array:3 [
              "identificador" => "bb0220"
              "etiqueta" => "Popovic et al&#46;&#44; 2019"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Extended <span class="elsevierStyleItalic">in vitro</span> culture of human embryos demonstrates the complex nature of diagnosing chromosomal mosaicism from a single trophectoderm biopsy"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:10 [
                            0 => "M&#46; Popovic"
                            1 => "L&#46; Dhaenens"
                            2 => "J&#46; Taelman"
                            3 => "A&#46; Dheedene"
                            4 => "M&#46; Bialecka"
                            5 => "P&#46; De Sutter"
                            6 => "Chuva de Sousa"
                            7 => "S&#46;M&#46; Lopes"
                            8 => "B&#46; Menten"
                            9 => "B&#46; Heindryckx"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1093/humrep/dez012"
                      "Revista" => array:7 [
                        "tituloSerie" => "Hum&#46; Reprod&#46;"
                        "fecha" => "2019"
                        "volumen" => "34"
                        "numero" => "4"
                        "paginaInicial" => "758"
                        "paginaFinal" => "769"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/30838420"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            44 => array:3 [
              "identificador" => "bb0225"
              "etiqueta" => "Reimundo et al&#46;&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Transferencia embrionaria &#250;nica&#58; estrategia clave para reducir el riesgo de embarazo m&#250;ltiple en reproducci&#243;n humana asistida"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [
                            0 => "P&#46; Reimundo"
                            1 => "J&#46; Guti&#233;rrez Romero"
                            2 => "T&#46; Rodr&#237;guez P&#233;rez"
                            3 => "E&#46; Veiga"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Adv&#46; Lab&#46; Med&#46;"
                        "fecha" => "2021"
                        "volumen" => "2"
                        "numero" => "2"
                        "paginaInicial" => "189"
                        "paginaFinal" => "198"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            45 => array:3 [
              "identificador" => "bb0230"
              "etiqueta" => "Rend&#243;n Abad&#44; 2018"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Cultivo de preembriones humanos en presencia de alta tensi&#243;n de ox&#237;geno&#58; resultados obst&#233;tricos y perinatales"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "M&#46; Rend&#243;n Abad"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Libro" => array:2 [
                        "fecha" => "2018"
                        "editorial" => "University of Valencia"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            46 => array:3 [
              "identificador" => "bb0235"
              "etiqueta" => "Rivron et al&#46;&#44; 2018"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Blastocyst-like structures generated solely from stem cells"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:10 [
                            0 => "N&#46;C&#46; Rivron"
                            1 => "J&#46; Frias-Aldeguer"
                            2 => "E&#46;J&#46; Vrij"
                            3 => "J&#46;C&#46; Boisset"
                            4 => "J&#46; Korving"
                            5 => "J&#46; Vivie"
                            6 => "R&#46;K&#46; Truckenmuller"
                            7 => "A&#46; van Oudenaarden"
                            8 => "C&#46;A&#46; van Blitterswijk"
                            9 => "N&#46; Geijsen"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46;"
                        "fecha" => "2018"
                        "volumen" => "557"
                        "numero" => "7703"
                        "paginaInicial" => "106"
                        "paginaFinal" => "111"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            47 => array:3 [
              "identificador" => "bb0240"
              "etiqueta" => "Rossant&#44; 2015"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Mouse and human blastocyst-derived stem cells&#58; vive les differences"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "J&#46; Rossant"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1242/dev.115451"
                      "Revista" => array:7 [
                        "tituloSerie" => "Development&#46;"
                        "fecha" => "2015"
                        "volumen" => "142"
                        "numero" => "1"
                        "paginaInicial" => "9"
                        "paginaFinal" => "12"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/25516964"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            48 => array:3 [
              "identificador" => "bb0245"
              "etiqueta" => "Rossant and Tam&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Opportunities and challenges with stem cell-based embryo models"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "J&#46; Rossant"
                            1 => "P&#46;P&#46; Tam"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Stem Cell Rep&#46;"
                        "fecha" => "2021"
                        "volumen" => "16"
                        "numero" => "5"
                        "paginaInicial" => "1031"
                        "paginaFinal" => "1038"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            49 => array:3 [
              "identificador" => "bb0250"
              "etiqueta" => "Shahbazi and Zernicka-Goetz&#44; 2018"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Deconstructing and reconstructing the mouse and human early embryo"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "M&#46; Shahbazi"
                            1 => "M&#46; Zernicka-Goetz"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/s41556-018-0144-x"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Cell Biol&#46;"
                        "fecha" => "2018"
                        "volumen" => "20"
                        "numero" => "8"
                        "paginaInicial" => "878"
                        "paginaFinal" => "887"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/30038253"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            50 => array:3 [
              "identificador" => "bb0255"
              "etiqueta" => "Shahbazi et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Self-organization of the human embryo in the absence of maternal tissues"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:14 [
                            0 => "M&#46;N&#46; Shahbazi"
                            1 => "A&#46; Jedrusik"
                            2 => "S&#46; Vuoristo"
                            3 => "G&#46; Recher"
                            4 => "A&#46; Hupalowska"
                            5 => "V&#46; Bolton"
                            6 => "N&#46;M&#46;E&#46; Fogarty"
                            7 => "A&#46; Campbell"
                            8 => "L&#46;G&#46; Devito"
                            9 => "D&#46; Ilic"
                            10 => "Y&#46; Khalaf"
                            11 => "K&#46;K&#46; Niakan"
                            12 => "S&#46; Fishel"
                            13 => "M&#46; Zernicka-Goetz"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/ncb3347"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Cell Biol&#46;"
                        "fecha" => "2016"
                        "volumen" => "18"
                        "numero" => "6"
                        "paginaInicial" => "700"
                        "paginaFinal" => "708"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27144686"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            51 => array:3 [
              "identificador" => "bb0260"
              "etiqueta" => "Shahbazi et al&#46;&#44; 2020"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Developmental potential of aneuploid human embryos cultured beyond implantation"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:12 [
                            0 => "M&#46;N&#46; Shahbazi"
                            1 => "T&#46; Wang"
                            2 => "X&#46; Tao"
                            3 => "B&#46;A&#46; Weatherbee"
                            4 => "L&#46; Sun"
                            5 => "Y&#46; Zhan"
                            6 => "L&#46; Keller"
                            7 => "G&#46;D&#46; Smith"
                            8 => "A&#46; Pellicer"
                            9 => "R&#46;T&#46; Scott"
                            10 => "E&#46; Seli"
                            11 => "M&#46; Zernicka-Goetz"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/s41467-019-13993-7"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Commun&#46;"
                        "fecha" => "2020"
                        "volumen" => "11"
                        "numero" => "1"
                        "paginaInicial" => "1"
                        "paginaFinal" => "15"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/31911652"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            52 => array:3 [
              "identificador" => "bb0265"
              "etiqueta" => "Shao et al&#46;&#44; 2017"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Self-organized amniogenesis by human pluripotent stem cells in a biomimetic implantation-like niche"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:10 [
                            0 => "Y&#46; Shao"
                            1 => "K&#46; Taniguchi"
                            2 => "K&#46; Gurdziel"
                            3 => "R&#46;F&#46; Townshend"
                            4 => "X&#46; Xue"
                            5 => "K&#46;M&#46; Yong"
                            6 => "J&#46; Sang"
                            7 => "J&#46;R&#46; Spence"
                            8 => "D&#46;L&#46; Gumucio"
                            9 => "J&#46; Fu"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/nmat4829"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Mater&#46;"
                        "fecha" => "2017"
                        "volumen" => "16"
                        "numero" => "4"
                        "paginaInicial" => "419"
                        "paginaFinal" => "425"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/27941807"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            53 => array:3 [
              "identificador" => "bb0270"
              "etiqueta" => "Sozen et al&#46;&#44; 2018"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Self-assembly of embryonic and two extra-embryonic stem cell types into gastrulating embryo-like structures"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:11 [
                            0 => "B&#46; Sozen"
                            1 => "G&#46; Amadei"
                            2 => "A&#46; Cox"
                            3 => "R&#46; Wang"
                            4 => "Czukiewska Ellen"
                            5 => "L&#46; Chappell"
                            6 => "T&#46; Voet"
                            7 => "G&#46; Michel"
                            8 => "N&#46; Jing"
                            9 => "D&#46;M&#46; Glover"
                            10 => "M&#46; Zernicka-Goetz"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1038/s41556-018-0147-7"
                      "Revista" => array:7 [
                        "tituloSerie" => "Nat&#46; Cell Biol&#46;"
                        "fecha" => "2018"
                        "volumen" => "20"
                        "numero" => "8"
                        "paginaInicial" => "979"
                        "paginaFinal" => "989"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/30038254"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            54 => array:3 [
              "identificador" => "bb0275"
              "etiqueta" => "Steptoe and Edwards&#44; 1978"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Birth after the reimplantation of a human embryo"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "P&#46;C&#46; Steptoe"
                            1 => "R&#46;G&#46; Edwards"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:5 [
                        "tituloSerie" => "Lancet&#46;"
                        "fecha" => "1978"
                        "volumen" => "312"
                        "numero" => "8085"
                        "paginaInicial" => "366"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            55 => array:3 [
              "identificador" => "bb0280"
              "etiqueta" => "Sunde et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Time to take human embryo culture seriously"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:8 [
                            0 => "A&#46; Sunde"
                            1 => "D&#46; Brison"
                            2 => "J&#46; Dumoulin"
                            3 => "J&#46; Harper"
                            4 => "K&#46; Lundin"
                            5 => "M&#46;C&#46; Magli"
                            6 => "E&#46; van den Abbeel"
                             …1
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1093/humrep/dew157"
                      "Revista" => array:7 [
                        "tituloSerie" => "Hum&#46; Reprod&#46;"
                        "fecha" => "2016"
                        "volumen" => "31"
                        "numero" => "10"
                        "paginaInicial" => "2174"
                        "paginaFinal" => "2182"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            56 => array:3 [
              "identificador" => "bb0285"
              "etiqueta" => "ThermoFisher Scientific&#44; 2014a"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Advanced DMEM&#47;F-12"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [ …1]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "WWW" => array:2 [
                        "link" => "https&#58;&#47;&#47;www&#46;thermofisher&#46;com&#47;order&#47;catalog&#47;product&#47;12634010"
                        "fecha" => "2014"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            57 => array:3 [
              "identificador" => "bb0290"
              "etiqueta" => "ThermoFisher Scientific&#44; 2014b"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Insulin-Transferrin-Selenium-Ethaolamine &#40;ITS-X&#41; &#40;100X&#41;"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [ …1]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "WWW" => array:2 [
                        "link" => "https&#58;&#47;&#47;www&#46;thermofisher&#46;com&#47;order&#47;catalog&#47;product&#47;51500056"
                        "fecha" => "2014"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            58 => array:3 [
              "identificador" => "bb0295"
              "etiqueta" => "ThermoFisher Scientific&#44; 2014c"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "AlbuMAX&#174; I and AlbuMAX&#174; II"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [ …1]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "WWW" => array:2 [
                        "link" => "https&#58;&#47;&#47;www&#46;thermofisher&#46;com&#47;document-connect&#47;document-connect&#46;html&#63;url&#61;https&#37;3A&#37;2F&#37;2Fassets&#46;thermofisher&#46;com&#37;2FTFS-Assets&#37;2FLSG&#37;2Fmanuals&#37;2F3117&#46;pdf"
                        "fecha" => "2014"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            59 => array:3 [
              "identificador" => "bb0300"
              "etiqueta" => "ThermoFisher Scientific&#44; 2020a"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Knockout&#8482; SR User Guide"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [ …1]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "WWW" => array:2 [
                        "link" => "https&#58;&#47;&#47;www&#46;thermofisher&#46;com&#47;order&#47;catalog&#47;product&#47;10828010&#63;SID&#61;srch-srp-10828010"
                        "fecha" => "2020"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            60 => array:3 [
              "identificador" => "bb0305"
              "etiqueta" => "ThermoFisher Scientific&#44; 2020b"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "GlutaMAX&#8482; Product Information Sheet"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [ …1]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "WWW" => array:2 [
                        "link" => "https&#58;&#47;&#47;assets&#46;fishersci&#46;com&#47;TFS-Assets&#47;LSG&#47;manuals&#47;GlutaMAX&#95;PI&#46;pdf&#63;&#95;ga&#61;2&#46;254523471&#46;695974732&#46;1645097875-1058616482&#46;1617892711&#38;&#95;gac&#61;1&#46;224585576&#46;1642763480&#46;CjwKCAiA0KmPBhBqEiwAJqKK49EYq0AMKeAS6erwSS&#95;dE8rEgLEcD9A1li5aUGnLL9hUWNG-7ItWeBoCRMcQAvD&#95;BwE"
                        "fecha" => "2020"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            61 => array:3 [
              "identificador" => "bb0310"
              "etiqueta" => "van den Brink et al&#46;&#44; 2014"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Symmetry breaking&#44; germ layer specification and axial organisation in aggregates of mouse embryonic stem cells"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:7 [ …7]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1242/dev.113001"
                      "Revista" => array:7 [
                        "tituloSerie" => "Development&#46;"
                        "fecha" => "2014"
                        "volumen" => "141"
                        "numero" => "22"
                        "paginaInicial" => "4231"
                        "paginaFinal" => "4242"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            62 => array:3 [
              "identificador" => "bb0315"
              "etiqueta" => "van der Valk et al&#46;&#44; 2018"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Fetal Bovine Serum &#40;FBS&#41;&#58; Past-Present-Future"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:17 [ …17]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Altex&#46;"
                        "fecha" => "2018"
                        "volumen" => "35"
                        "numero" => "1"
                        "paginaInicial" => "1"
                        "paginaFinal" => "20"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            63 => array:3 [
              "identificador" => "bb0320"
              "etiqueta" => "van Montfoort et al&#46;&#44; 2020"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Reduced oxygen concentration during human IVF culture improves embryo utilization and cumulative pregnancy rates per cycle"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:7 [ …7]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1093/hropen/hoz036"
                      "Revista" => array:6 [
                        "tituloSerie" => "Hum&#46; Reprod&#46; Open&#46;"
                        "fecha" => "2020"
                        "volumen" => "2020"
                        "numero" => "1"
                        "paginaInicial" => "hoz036"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            64 => array:3 [
              "identificador" => "bb0325"
              "etiqueta" => "Wale and Gardner&#44; 2010"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Time-lapse analysis of mouse embryo development in oxygen gradients"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.rbmo.2010.04.028"
                      "Revista" => array:7 [
                        "tituloSerie" => "Reprod&#46; BioMed&#46; Online"
                        "fecha" => "2010"
                        "volumen" => "21"
                        "numero" => "3"
                        "paginaInicial" => "402"
                        "paginaFinal" => "410"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            65 => array:3 [
              "identificador" => "bb0330"
              "etiqueta" => "Wang et al&#46;&#44; 2002"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Vitamin C and vitamin E supplementation reduce oxidative stress&#8211;induced embryo toxicity and improve the blastocyst development rate"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:6 [ …6]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/s0015-0282(02)04236-x"
                      "Revista" => array:7 [
                        "tituloSerie" => "Fertil&#46; Steril&#46;"
                        "fecha" => "2002"
                        "volumen" => "78"
                        "numero" => "6"
                        "paginaInicial" => "1272"
                        "paginaFinal" => "1277"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            66 => array:3 [
              "identificador" => "bb0335"
              "etiqueta" => "Warnock&#44; 1984"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Report of the Committee of Inquiry into Human Fertilization and Embryology"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [ …1]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Libro" => array:3 [
                        "fecha" => "1984"
                        "editorial" => "Her Majesty&#8217;s Stationery Office &#40;HMSO&#41;"
                        "editorialLocalizacion" => "London"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            67 => array:3 [
              "identificador" => "bb0340"
              "etiqueta" => "Weimar et al&#46;&#44; 2013"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "In-vitro model systems for the study of human embryo-endometrium interactions"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:5 [ …5]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.rbmo.2013.08.002"
                      "Revista" => array:6 [
                        "tituloSerie" => "Reprod&#46; BioMed&#46; Online"
                        "fecha" => "2013"
                        "volumen" => "27"
                        "numero" => "5"
                        "paginaInicial" => "461&#46;-476"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            68 => array:3 [
              "identificador" => "bb0345"
              "etiqueta" => "Williams and Johnson&#44; 2020"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Adapting the 14-day rule for embryo research to encompass evolving technologies"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.rbms.2019.12.002"
                      "Revista" => array:6 [
                        "tituloSerie" => "Reprod&#46; Biomed&#46; Soc&#46; Online&#46;"
                        "fecha" => "2020"
                        "volumen" => "10"
                        "paginaInicial" => "1"
                        "paginaFinal" => "9"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            69 => array:3 [
              "identificador" => "bb0350"
              "etiqueta" => "Wu and Morris Jr&#44; 1998"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Arginine metabolism&#58; nitric oxide and beyond"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1042/bj3360001"
                      "Revista" => array:7 [
                        "tituloSerie" => "Biochem&#46; J&#46;"
                        "fecha" => "1998"
                        "volumen" => "336"
                        "numero" => "1"
                        "paginaInicial" => "1"
                        "paginaFinal" => "17"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            70 => array:3 [
              "identificador" => "bb0355"
              "etiqueta" => "Zander-Fox et al&#46;&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Albumin used in human IVF contain different levels of lipids and modify embryo and fetal growth in a mouse model"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [ …3]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1007/s10815-021-02255-5"
                      "Revista" => array:6 [
                        "tituloSerie" => "J&#46; Assist&#46; Reprod&#46; Genet&#46;"
                        "fecha" => "2021"
                        "volumen" => "38"
                        "paginaInicial" => "2371"
                        "paginaFinal" => "2381"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            71 => array:3 [
              "identificador" => "bb0360"
              "etiqueta" => "Zernicka-Goetz et al&#46;&#44; 2016"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Culture of Human Embryos through Implantation Stages <span class="elsevierStyleItalic">in Vitro</span>"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [ …4]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Libro" => array:3 [
                        "fecha" => "2016"
                        "paginaInicial" => "1"
                        "paginaFinal" => "7"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            72 => array:3 [
              "identificador" => "bb0365"
              "etiqueta" => "Zhai et al&#46;&#44; 2021"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Human embryonic development&#58; from peri-implantation to gastrulation"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:4 [ …4]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/j.tcb.2021.07.008"
                      "Revista" => array:7 [
                        "tituloSerie" => "Trends Cell Biol&#46;"
                        "fecha" => "2021"
                        "volumen" => "32"
                        "numero" => "1"
                        "paginaInicial" => "18"
                        "paginaFinal" => "29"
                        "link" => array:1 [
                          0 => array:2 [ …2]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            73 => array:3 [
              "identificador" => "bb0370"
              "etiqueta" => "Zheng et al&#46;&#44; 2019"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Controlled modelling of human epiblast and amnion development using stem cells"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:11 [ …11]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Nat&#46;"
                        "fecha" => "2019"
                        "volumen" => "573"
                        "numero" => "7774"
                        "paginaInicial" => "421"
                        "paginaFinal" => "425"
                      ]
                    ]
                  ]
                ]
              ]
            ]
          ]
        ]
      ]
    ]
  ]
  "idiomaDefecto" => "en"
  "url" => "/23409320/0000000900000003/v1_202210200635/S2340932022000068/v1_202210200635/en/main.assets"
  "Apartado" => array:4 [
    "identificador" => "93980"
    "tipo" => "SECCION"
    "en" => array:2 [
      "titulo" => "Revisiones"
      "idiomaDefecto" => true
    ]
    "idiomaDefecto" => "en"
  ]
  "PDF" => "https://static.elsevier.es/multimedia/23409320/0000000900000003/v1_202210200635/S2340932022000068/v1_202210200635/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/"
  "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S2340932022000068?idApp=UINPBA00004N"
]

Article information

ISSN: 23409320
Original language: English

DOI:10.1016/j.medre.2022.100118

The statistics are updated each day

Year/Month	Html	Pdf	Total

2024 November	14	4	18
2024 October	158	24	182
2024 September	193	34	227
2024 August	120	26	146
2024 July	39	5	44
2023 March	1	0	1
2022 November	2	2	4
2022 October	16	5	21

Follow us:

Statistics

Subscribe to our newsletter