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体外诱导人脐带间充质干细胞向卵母细胞样细胞分化的性别依赖性潜能。

Gender depended potentiality of differentiation of human umbilical cord mesenchymal stem cells into oocyte-Like cells in vitro.

机构信息

College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University, Yangling, Shaanxi, China.

出版信息

Cell Biochem Funct. 2013 Jul;31(5):365-73. doi: 10.1002/cbf.2981. Epub 2013 May 9.

Abstract

Recent studies have demonstrated that germ-like cells could be differentiated from human umbilical cord mesenchymal stem cells (hUC-MSCs) in vitro. Whether the sexuality of hUC-MSCs affects the formation efficiency of germ-like cells derived from hUC-MSCs is still unclear. To clearly test the formation efficiency of oocyte-like cells from male and female hUC-MSCs, obtained hUC-MSCs were induced by 20% follicular fluid (FF) according to the method that has been proved by our previous studies. Results showed that hUC-MSCs differentiated into oocyte-like structures and expressed germ cell makers. It was noted that the presence of advanced oocyte-like cells in male hUC-MSCs (m-hUC-MSCs) was similar as that in female hUC-MSCs (f-hUC-MSCs); however, the expression of germ cell's specific markers in m-hUC-MSCs was delayed compared with that in f-hUC-MSCs. In addition, immunofluorescence analysis demonstrated that germ cell-specific markers, Oct4, Vasa, Dazl, ZP2, ZP3 and Stra8, were expressed on the 14th day after induction in both f-hUC-MSCs and m-hUC-MSCs. However, the size of oocyte-like cells from f-hUC-MSCs was larger than that in m-hUC-MSCs. The level of secreted oestradiol was significantly higher in f-hUC-MSCs than m-hUC-MSCs. We sought to determine whether critical germ cell's transcription factor-Figlα will promote the development of oocyte-like cells. Some germ cell-specific markers were increased when exogenous Figlα was transfected into hUC-MSCs. This process implied that germ-like cells might be produced by over-expression of exogenous germ cell-specific gene, and this process was similar as that in production of germ cells in induced pluripotent stem cells (iPSCs). Finally, to verify the feasibility that hUC-MSCs differentiate into germ cells, hUC-MSCs were transplanted into seminiferous tubules and kidney capsule of mouse, respectively, and we found the transplanted cells differentiated into germ-like cells in recipient's seminiferous tubules and kidney capsule. This study will provide a simple model to study mammalian germ cell specification using hUC-MSCs in vitro.

摘要

最近的研究表明,类 germ 细胞可在体外从人脐带间充质干细胞(hUC-MSCs)中分化而来。hUC-MSCs 的性别是否影响源自 hUC-MSCs 的类 germ 细胞的形成效率尚不清楚。为了清楚地测试来自男性和女性 hUC-MSCs 的卵母细胞样细胞的形成效率,根据我们之前研究证明的方法,用 20%卵泡液(FF)诱导获得的 hUC-MSCs。结果表明,hUC-MSCs 分化为卵母细胞样结构并表达 germ 细胞标记物。值得注意的是,男性 hUC-MSCs(m-hUC-MSCs)中晚期卵母细胞样细胞的存在与女性 hUC-MSCs(f-hUC-MSCs)相似;然而,m-hUC-MSCs 中 germ 细胞特异性标记物的表达比 f-hUC-MSCs 延迟。此外,免疫荧光分析表明,在诱导后的第 14 天,f-hUC-MSCs 和 m-hUC-MSCs 中均表达 germ 细胞特异性标记物 Oct4、Vasa、Dazl、ZP2、ZP3 和 Stra8。然而,f-hUC-MSCs 来源的卵母细胞样细胞的大小大于 m-hUC-MSCs。f-hUC-MSCs 分泌的雌二醇水平明显高于 m-hUC-MSCs。我们试图确定关键的 germ 细胞转录因子-Figlα 是否会促进卵母细胞样细胞的发育。当外源性 Figlα 转染到 hUC-MSCs 中时,一些 germ 细胞特异性标记物增加。这个过程意味着 germ 样细胞可能是通过过表达外源性 germ 细胞特异性基因产生的,这个过程类似于诱导多能干细胞(iPSCs)中 germ 细胞的产生。最后,为了验证 hUC-MSCs 分化为 germ 细胞的可行性,我们分别将 hUC-MSCs 移植到小鼠的生精小管和肾包膜中,发现移植细胞在受者的生精小管和肾包膜中分化为 germ 样细胞。这项研究将提供一个使用 hUC-MSCs 体外研究哺乳动物 germ 细胞特化的简单模型。

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