传代培养人胚胎生殖干细胞的生物学鉴定

Biological identification of passage cultured human embryonic germ stem cells

目的：通过对传代培养至第5代细胞分子标记物的表达分析,鉴定传代培养的细胞为人胚胎生殖干细胞(embrgonic germ stem cells,EGCs)。方法：取5～10周人胚胎的生殖腺嵴和肠背系膜,进行组织块体外培养人EGCs,挑选传代培养至第5代生长良好的细胞,采用免疫细胞化学法分别检测SSEA-1、SSEA-3、Oct-4、hTERT的表达。结果：组织块体外培养得到的细胞,经免疫细胞化学法显示的单个细胞或细胞集落,均阳性表达SSEA- 1、SSEA-3、Oct-4、hTERT。结论：组织块体外传代培养第5代所获得的细胞,为未分化的人EGCs。

Objective： To identify the passage cultured cells as human embryonic germ stem cells by detecting the expression of molecular markers in the serial subcultured cells. Methods： Primary human embryonic germ stem cells were obtained by tissue culturing of embryonic genital ridges and mesenteries from the embryos aged 5 to 10 weeks and then were passaged 4 times. Immunocytochemical staining for four specific markers including SSEA-1, SSEA-3, Oct-4 and hTERT was applied to ascertain whether these subcultured cells were human embryonic germ cells. Results： Immunocytochemistry showed that the single cells and the colonies of the fifth generation cells expressed 4 kinds of molecular markers. Conclusion： The fifth generation cells obtained through tissue culturing in vitro are undifferentiated human embryonic germ stem cells （EGCs）.