人卵母细胞样细胞体内分化模型的建立

Generation of human oocyte-like cell differentiation in vivo

摘要:

干细胞通过诱导培养，在体外能够分化为卵母细胞样细胞 (Oocyte-like cell, OLC)，将其置于体内环境能够有效地改善OLC的质量和发育能力。通过检测猪卵泡液中激素和Bmp 15蛋白的含量，选取了中等卵泡的卵泡液对人羊水干细胞进行体外诱导培养，10 d后，经实时定量PCR检测发现，早期生殖细胞样细胞团高表达生殖基因oct4和重新甲基化转移酶基因dnmt3b。将这些细胞团用猪卵泡膜包裹后形成移植物，移植到小鼠肾被膜下。1个月后取出移植物，发现移植物内的细胞在形态上，不仅与正常的卵母细胞极为相似，而且还表达生殖细胞和卵母细胞特异标记基因 (oct4、nanog、stella、ifitm3、dazl、nanos3、bmp15和gdf9)。证明技术体系能够有效改善OLC的形成和发育能力。

关键词:人羊水干细胞，卵母细胞样细胞，骨形成蛋白15，卵泡液，分化

Abstract:

Oocyte-like cells (OLC) can be generated by stem cells after the induction and differentiation in vitro, and maturated when transplanted in vivo to improve the development potential. Human amniotic fluid stem cells (hAFSC) were cultured for 10 days in porcine follicle fluid (pFF) that was extracted from the medium follicle with high levels of hormones and Bmp 15 protein. After the induction, the cell aggregates showed the germ cell-like cells and produced the germ cell marker oct4, and triggered epigenetic changes with high expression of methylation transferase gene dnmt3b. The cell aggregates were packaged into porcine theca folliculi to form grafts, which were then transplanted into mouse renal capsule. After one month of transplantation, the morphology of OLC from a graft was not only similar to oocytes, but also expressed the germ cells markers (oct4, nanog, stella, ifitm3, dazl, nanos3, bmp15, and gdf9). The results demonstrate that the in vivo differentiation model was useful for OLC development.

Key words:human amniotic fluid stem cell, oocyte-like cell, Bmp15, follicle fluid, differentiation