小鼠睾丸支持细胞的分离培养与生物学研究

目的]分离得到高纯度的小鼠睾丸支持细胞(Sertoli cell),用以研究它对精原干细胞(SSCs)生长的影响。方法]取鼠龄3~7 d的雄性ICR小鼠睾丸,采用胶原酶、透明质酸酶、胰蛋白酶和DNA酶消化分离Sertoli细胞及SSCs,经多次差异离心贴壁及低渗处理法进行纯化,用Sertoli细胞作饲养层观察SSCs的生长情况。结果]Sertoli细胞纯度达90%,且以Sertoli细胞作饲养层培养SSCs,与对照相比,能明显促进SSCs的增殖。结论]四酶消化低渗处理法可获得高纯度的Sertoli细胞,且Sertoli细胞能明显促进SSCs的增殖。

Culture and Biological Studies on Sertoli Cells Isolated from Mouse Testis

Sertoli cells from mouse testis were isolated and cultured to study the effect of Sertoli cells feeders on mouse spermatogonial stem cells in vitro .Sertoh ceils and spermatogonial stem ceils were isolated from 3- 7 d ICR mouse and subjected to sequential enzymatic treatment. Sertuli cells were treated with Tris-HCl to remove spermatogenic cells and high purity sertoli cells were obtained. Sertoli cells treated with mitomycin C were used as feeder cells, to study the effect on mouse spermatogonial stem cells. After a week of culture, compared with control, there were more number of spermatogonial stem cells remained. More than 90% of the Sertoli cells were isolated and cultured. Sertoli cells can promote proliferation of spermatogonial stem cells obviously. More Sertoli cells were obtained through this experiment method with high purification. Sertoli cells can be used as feeders not only to promote survival but also renewal of spermatogonial stem cells.