SummaryAndrogenetic embryonic stem (AgES) cells offer a possible tool for patient-specific pluripotent stem cells that will benefit genomic imprinting studies and clinic applications. However, the difficulty in producing androgenetic embryos and the unbalanced expression of imprinted genes make the therapeutic applicability of AgES cells uncertain. In this study, we produced androgenetic embryos by injecting two sperm into an enucleated metaphase II (MII) oocyte. By this method, 88.48% of oocytes survived after injection, and 20.24% of these developed to the blastocyst stage. We successfully generated AgES cell lines from the androgenetic embryos and assayed the expression of imprinted genes in the cell lines. We found that the morphological characteristics of AgES cells were similar to that of fertilized embryonic stem cells (fES), such as expression of key pluripotent markers, and generation of cell derivatives representing all three germ layers following in vivo and in vitro differentiation. Furthermore, activation of paternal imprinted genes was detected, H19, ASC12 and Tss3 in AgES cell activation levels were lower while other examined genes showed no significant difference to that of fES cells. Interestingly, among examined maternal imprinted genes, only Mest and Igf2 were significantly increased, while levels of other detected genes were no different to that of fES cells. These results demonstrated that activation of some paternal imprinted genes, as well as recovery of maternal imprinted genes, was present in AgES cells. We differentiated AgES cells into a beating embryoid body in vitro, and discovered that the AgES cells did not show significant higher efficiency in myocardial differentiation potential.

中文翻译：

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双精子注射建立小鼠雄激素胚胎干细胞并分化为跳动胚体

摘要雄激素胚胎干细胞 (AgES) 为患者特异性多能干细胞提供了一种可能的工具，这将有利于基因组印记研究和临床应用。然而，雄激素胚胎的产生困难和印迹基因的不平衡表达使得AgES细胞的治疗适用性不确定。在这项研究中，我们通过将两个精子注射到去核中期 II (MII) 卵母细胞中来产生雄激素胚胎。通过这种方法，88.48%的卵母细胞在注射后成活，其中20.24%的卵母细胞发育至囊胚期。我们成功地从雄激素胚胎中产生了 AgES 细胞系，并分析了印迹基因在细胞系中的表达。我们发现AgES细胞的形态特征与受精胚胎干细胞（fES）相似，体内和体外差异化。此外，检测到父系印记基因的激活，H19,ASC12和TS3AgES 细胞活化水平较低，而其他检查的基因与 fES 细胞没有显着差异。有趣的是，在检查过的母体印记基因中，只有梅斯特和胰岛素样生长因子2显着增加，而其他检测到的基因水平与 fES 细胞没有差异。这些结果表明，一些父系印记基因的激活以及母体印记基因的恢复存在于 AgES 细胞中。我们将 AgES 细胞分化成跳动的胚状体体外, 并发现 AgES 细胞在心肌分化潜能方面没有表现出显着更高的效率。