小鼠睾丸间质细胞的分离培养

Isolation and Culture of Mouse Leydig Cells

寻求一种简单有效的方法分离纯化睾丸间质细胞并寻求较适宜的培养条件。采用机械撕碎、密度梯度离心等分离、纯化细胞,设对照组寻求较适宜的培养条件。刚分离的细胞经苔盼蓝染色,存活率>90%;体外培养的细胞形态完整、增殖速度快、贴壁生长状态良好。用17-α羟化酶和用雄激素结合蛋白RT-PCR扩增进行纯度鉴定,前者出现特异片段,后者未出现。该方法简单、经济、快速、有效,适合睾丸间质体外分离培养。

To obtain a simple and effective method and eligible condition to isolate and culture mouse Leydig cells. The testes of mouse were torn up with forceps, isolated and purified through density gradient centrifugation. Control groups were set up to seek fairly eligible growing condition. The characteristic of cells were identified through cytological observation and Trypanblau staining, their survival rate〉90%. The results showed that the Leydig cells retained complete shape, good biologic activity, growing up quickly in adherence. The amplication of CYP17 were observed in agarose gel by electrophoresis and the ABP without gel electrophoresis band. The method is simple, rapid, effective and suited to isolate and culture Leydig cells in vitro.