Reproductive Medical Center, Department of Obstetrics and Gynecology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Gynecol Endocrinol. 2012 Jan;28(1):25-8. doi: 10.3109/09513590.2011.588748. Epub 2011 Jul 23.
Recently, human embryonic stem cells (hESCs) of some genetic diseases have been established, but little research has been done on polycystic ovary syndrome (PCOS)-derived hESCs. The establishment of PCOS-derived hESCs provides a biological basis for exploring the pathogenesis, gene mapping and gene therapy of PCOS.
Discarded fresh embryos were collected and cultured until the blastocyst stage, and then inner cell masses (ICM) were isolated by mechanical methods and incubated in the mixed feeder layer containing human stem cell medium. hESCs were identified whether to maintain normal karyotype and pluripotency by alkaline phosphates (AKP), stage-specific embryonic antigen-4 (SSEA-4), NANOG, SOX2 and TRA-1-60, octamer binding protein 4(OCT-4), and in vivo and in vitro differentiation.
Of the 11 passaged ICM, nine showed adherent growths within 48 h with an adherence rate of 81.8% (9/11). Five PCOS-derived hESCs were established and all of them have the characteristics of pluripotent differentiation. One was from 2PN embryo which was retarded in the cleavage stage, one was from 1PN embryo and others were from 0PN embryo. They were named p-hES-1, p-hES-2, p-hES-3, p-hES-4, p-hES-5, respectively.
We provide biological models for studying the pathogenesiss of PCOS.
最近，一些遗传疾病的人类胚胎干细胞（hESC）已经建立，但对多囊卵巢综合征（PCOS）来源的 hESC 研究甚少。PCOS 来源的 hESC 的建立为探索 PCOS 的发病机制、基因定位和基因治疗提供了生物学基础。
收集废弃的新鲜胚胎并培养至囊胚阶段，然后通过机械方法分离内细胞团（ICM），并在含有人干细胞培养基的混合饲养层中孵育。通过碱性磷酸酶（AKP）、阶段特异性胚胎抗原-4（SSEA-4）、NANOG、SOX2 和 TRA-1-60、八聚体结合蛋白 4（OCT-4），以及体内和体外分化来鉴定 hESC 是否保持正常核型和多能性。
在传代的 11 个 ICM 中，有 9 个在 48 小时内呈贴壁生长，贴壁率为 81.8%（9/11）。建立了 5 株 PCOS 来源的 hESC，均具有多能分化的特征。其中一株来自于卵裂期发育迟缓的 2PN 胚胎，一株来自于 1PN 胚胎，另外三株来自于 0PN 胚胎。它们分别被命名为 p-hES-1、p-hES-2、p-hES-3、p-hES-4、p-hES-5。
我们为研究 PCOS 的发病机制提供了生物学模型。
