大鼠睾丸支持细胞的原代培养与鉴定

THE PRIMARY CULTURE AND IDENTIFICATION OF RAT SERTOLI CELLS

目的采用改良方法对大鼠睾丸支持细胞进行分离、纯化和原代培养,为得到更高纯度和稳定的支持细胞原代培养体系,并建立一种简单、易行的支持细胞鉴定方法。方法采用酶消化法分离大鼠睾丸支持细胞,用Tris-HCl处理后除去生精细胞,实现进一步纯化。并用Feulgen染色法鉴定支持细胞,对支持细胞进行形态学观察。结果培养的支持细胞纯度达到95%,每个睾丸可获取支持细胞约107个,Feulgen染色后明显可见支持细胞核内的双极小体。结论酶消化法分离和Feulgen染色法鉴定大鼠支持细胞简单易行,且细胞纯度较高。

Objective We used modified method to separate and culture Sertoli cells in order to set up a stable primary culture system of Sertoli cells. Besides, a simple but feasible way was founded to identify Sertoli cells. Methods We used enzymatic digestion to separate Sertoli cells, and treated with Tris-HCl to remove spermatogenic cells. Identified the Sertoli ceils with Feulgen staining, then viewed the morphology of cultured Sertoli cells under the microscope. Results The purity of Sertoli cells came to 95 percent, the total number of Sertoli cells we got from one testicle was up to 10^7 , bipolar corpuscula in nucleus was clearly observed after Feulgen staining. Conclusion The method of enzymatic digestion and Feulgen staining to separate and identify Sertoli cells is simple and feasible with high purity.