Reproductive Medical Center, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
J Cell Biochem. 2012 Dec;113(12):3835-42. doi: 10.1002/jcb.24297.
In recent years, applications of stem cells have already involved in all domains of life science and biomedicine. People try to establish human embryonic stem cell lines (hESCs) in order to carry out hESC-related studies. In this study, we explored what embryos are conducive to the establishment of hESCs. The discarded embryos from in vitro fertilization-embryo transfer (IVF-ET) cycles were sequentially incubated into blastocysts, and then the inner cell mass (ICM) was isolated and incubated in the mixed feeder layer. The cell lines which underwent serial passage were identified. After a total of 1,725 discarded embryos from 754 patients were incubated, 448 blastocysts were formed with 123 high-quality blastocysts. The blastulation rate was significantly higher in the discarded embryos with non-pronucleus (0PN) or 1PN than in the discarded embryos with 2PN or ≥3PN. The blastulation rate of the D3 embryos with 7-9 blastomeres was higher. Among the originally incubated 389 ICMs, 22 hESCs with normal karyotype were established, and identified to be ESCs. Therefore, in establishing hESCs with discarded embryos, D(3) 0PN or 1PN embryos with 7-9 blastomeres should be first selected, because they can improve high-quality blastulation rate which can increase the efficiency of hESC establishment.
近年来，干细胞的应用已经涉及生命科学和生物医学的各个领域。人们试图建立人类胚胎干细胞系（hESC），以进行与 hESC 相关的研究。在这项研究中，我们探讨了哪些胚胎有利于建立 hESC。我们将体外受精-胚胎移植（IVF-ET）周期中废弃的胚胎依次孵育为囊胚，然后分离内细胞团（ICM）并在混合饲养层中孵育。对经过连续传代的细胞系进行了鉴定。在孵育了 754 名患者的 1725 个废弃胚胎后，形成了 448 个囊胚，其中 123 个是高质量的囊胚。无原核（0PN）或 1PN 的废弃胚胎的囊胚形成率明显高于 2PN 或≥3PN 的废弃胚胎。具有 7-9 个卵裂球的 D3 胚胎的囊胚形成率更高。在最初孵育的 389 个 ICM 中，建立了 22 株具有正常核型的 hESC，并鉴定为 ESCs。因此，在利用废弃胚胎建立 hESC 时，应首先选择 D（3）0PN 或 1PN 的具有 7-9 个卵裂球的胚胎，因为它们可以提高高质量的囊胚形成率，从而提高 hESC 建立的效率。
