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首页> 外文期刊>The Journal of Reproduction and Development >Epiblast Cell Number and Primary Embryonic Stem Cell Colony Generation Are Increased by Culture of Cleavage Stage Embryos in Insulin
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Epiblast Cell Number and Primary Embryonic Stem Cell Colony Generation Are Increased by Culture of Cleavage Stage Embryos in Insulin

机译：胰岛素的卵裂期胚胎培养可增加成胚细胞数和原代胚胎干细胞集落的产生

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摘要

Human embryos for hESC derivation are often donated at the cleavage stage and of reduced quality. Poor quality embryos have lower efficiency for hESC derivation. However, cleavage stage mouse embryos develop into higher quality expanded blastocysts if they are cultured with insulin, suggesting that this approach could be used to improve hESC derivation from poor quality cleavage stage embryos. The present study used a mouse model to examine this approach. In particular we examined the effect of insulin on the number of epiblast cells in blastocysts on days 4, 5 and 6 using Oct4 and Nanog co-expression. Second we examined the effect of insulin on the frequency with which outgrowths can be derived from these. Finally, we tested whether prior culture in the presence of insulin results in blastocysts with increased capacity to generate ESC colonies. Culture of cleavage stage embryos with insulin increased the number of Oct4 and Nanog positive cells in blastocysts at all time points examined. Prior culture with insulin had no effect on outgrowths generated from blastocysts plated on days 4 or 5. However, insulin treatment of blastocysts plated on day 6 resulted in increased numbers of outgrowths with larger epiblasts compared with controls. 13% of insulin treated day 6 blastocysts produced primary ESC colonies compared with 6% of controls. In conclusion, treatment with insulin can improve epiblast cell number in mice leading to an increase with which primary ESC colonies can be generated and may improve hESC isolation from reduced quality embryos donated at the cleavage stage.
机译：用于hESC衍生的人类胚胎通常在卵裂期捐赠，且质量下降。劣质胚胎的hESC衍生效率较低。但是，如果将卵裂期小鼠胚胎与胰岛素一起培养，它们会发育成更高质量的膨胀胚泡，这表明该方法可用于改善从质量差的卵裂期胚胎中衍生的hESC。本研究使用小鼠模型来检查这种方法。特别是，我们使用Oct4和Nanog共表达检查了胰岛素在第4、5和6天对囊胚中上皮细胞数量的影响。其次，我们检查了胰岛素对这些衍生结果产生频率的影响。最后，我们测试了在胰岛素存在下的先前培养是否会导致胚泡的生成ESC集落的能力增加。在所有检查的时间点，用胰岛素培养卵裂期的胚胎会增加胚泡中Oct4和Nanog阳性细胞的数量。事先用胰岛素培养对第4天或第5天接种的囊胚产生的增生没有影响。但是，与第6天相比，对第6天接种的囊胚进行胰岛素治疗会导致增生的数量增加。胰岛素处理的第6天囊胚中有13％产生了原发性ESC菌落，而对照组则为6％。总之，用胰岛素治疗可以改善小鼠的表皮细胞数目，从而增加原代ESC菌落的产生，并可以改善hESC从卵裂期捐赠的质量下降的胚胎中的分离。

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