叙利亚地鼠原始生殖细胞体外培养方法的初步研究

An Initial Investigation on the In Vitro Culture System of Primordial Germ Cells in Golden Hamsters

为建立叙利亚地鼠原始生殖细胞的体外培养方法,在体外无菌分离孕龄10.5 d的叙利亚地鼠胚胎的生殖嵴,采用酶机械分离方法获取原始生殖细胞,在条件培养基中培养于饲养层细胞上,然后用酶组织化学染色方法鉴定碱性磷酸酶(ALP)的活性。去除分化抑制因素并诱导原始生殖细胞体外分化后,观察拟胚体和分化细胞的产生。实验结果显示孕龄10.5 d的地鼠胚胎生殖嵴,用酶机械分离方法能成功地分离出具有发育多能性的原始生殖细胞,ALP染色鉴定呈强阳性,体外诱导分化后可观察到3种胚层来源的细胞。结果提示该细胞体外培养体系的建立,为生物医学工程学提供了一种新的细胞来源。

To establish the in vitro culture system of primordial germ cells （PGCs） of golden hamsters, PGCs of hamster were isolated from genital ridge of embryos at 10. 5＇h dpc （day post coitum）, obtained by enzymemechanical method, and cultured on feeder cells. Then the PGCs were identified by alkaline phosphatase （ALP） activity staining. In order to induce the PGCs to differentiate in vitro, we removed the differential inhibition factors in the conditioned medium and observed the formation of embryoids and differentiated cells from three layers. The result showed that the pluripotent primordial germ cells could be successfully obtained from the golden hamsters at 10. 5th dpc by the enzyme-mechanical method and that PGCs were identified by both their strong positive reaction in ALP activity staining test and their differentiation into three-layer derived cells in vitro. The result suggests that the establishment of in vitro PGCs culture system of golden hamsters will provide new cell source for biomedical engineering.