大鼠睾丸支持细胞的分离纯化与鉴定

采用Ⅴ型胶原酶单次振荡消化法和低渗处理,及差速贴壁和免疫组化染色法,对大鼠睾丸支持细胞进行分离、纯化和鉴定。结果表明,该分离纯化方法对睾丸支持细胞损伤较小,获得的细胞活性较高,原代细胞存活率为85%,且无较大细胞团,接种后增殖迅速,并在第8天达到最高峰;细胞产率为6.5×105/g;绝大多数细胞呈F as-L阳性,细胞纯度可达95%;细胞核仁清楚,核仁周围可见着色较深的卫星核小体。

Isolation,purification and identification of testis Sertoli cells in prepubertal rats

Rats Sertoli cells were isolated,purified and identified by one-time digestion with collagenase V,differential adhesion,low osmotic buffer treatment and immunohistochemical staining.Results demonstrated that this isolation and purificatio method of Sertoli cells did little harm to the cells.The primarily isolated Sertoli cells had good cell viability;cell survival rate reached 85% and there was no big cell corps;Planted in flasks,cell proliferated rapidly and reached the proliferation peak at day 8 postcultured.Cell yield was 6.5×10⁵ cell/g;Most acquired cells were Fas-L positively stained,and cell purity could reach 95%;the nucleolus of the cells was clear,and there were satellite nucleosomes around the nucleolus.