改良的大鼠睾丸支持细胞分离培养方法

The Modified Isolation and Culture Method of Rat Sertoli Cells

目的 简化睾丸支持细胞的分离方法 ,提高细胞产量。方法 采用 0 .2 5 %胰酶、0 .0 5 %胶原酶序贯两步消化法消化 ,细胞悬液在 37°C、5 % CO2 培养箱内孵育培养 4 8h,观察产量和细胞功能。结果 改良的分离方法所获得的睾丸支持细胞占细胞总数的 90 %以上 ,大大提高了支持细胞的获取量 ,细胞 Fas配体 (Fas- L )表达功能正常。结论 改良的分离培养方法简化了传统的分离培养方法 。

Objective To simplify the procedures of isolation of rat Sertoli cells and increase the yield of Sertoli cells cultured. Methods Decapsulated rat testis from 16 22 day old SD rats were digested by sequential two step digestion using 0.25% pancreatin, 0.05% collagenase in order, and then the cell suspension was cultured in incubator under the condition of 37℃ and 5% CO 2 for 48 h. The yields were observed and Fas Ligand(Fas L) was tested by immunohistochemical method. Results By the use of this modified method, the yielded sertoli cells accounted for 90% of all the cells harvested. High level expression of Fas L was detected. Conclusion This modified method increases the yield of Sertoli cells and the procedures of isolation and culture are greatly simplified.