犊牛睾丸支持细胞的原代培养与鉴定

寻求简便经济的方法分离纯化犊牛睾丸支持细胞,通过不同的方法对支持细胞进行鉴定。采用胶原酶和胰蛋白酶次第消化分离犊牛睾丸支持细胞,5 h后换液,24 h后用Tris-HCl处理除去精原细胞,实现进一步纯化。用Feulgen染色法鉴定支持细胞,用SABC染色法检测支持细胞上Fas-L的表达。结果培养的支持细胞纯度达到90%以上。Feulgen染色后明显可见支持细胞核内的双极小体。SABC染色法检测支持细胞为阳性。建立了培养犊牛睾丸支持细胞的方法,Feulgen染色和Fas-L检测是鉴定支持细胞的有效方法。

Priory Culture Identification of testis Sertoli cells in calf

To search for a simple and economical method to separate and purify calf testis sertoli cells and to identify the sertoli cells with different method,the method of collagenase and typin was sequentially used to separate sertoli cells.After 5 hours the culture medium was replenished and after 24 hours the treatment with Tris-HC was done to remove spermatogenic cells.The sertoli cells were identified with Feulgen staining.The expression of FAS-L was examined by SABC staining.The result indicated that the purity of sertoli cells accounted for over 90% of all cells harvested.Bipolar corpuscula in nucleus was clearly observed after Feulgen staining.Sertoli cells expressed FAS-L.The conclusion was the method of isolation and culture of sertoli cells from calf was established.Feulgen staining and SABC staining were useful method for identification of the sertoli cells.