大鼠睾丸支持细胞的分离纯化与鉴定

目的　简化大鼠睾丸支持细胞的分离纯化方法 ,通过不同方法对Sertoli细胞进行形态学观察鉴定。方法　取鼠龄 16～ 2 2天的雄性Wistar大鼠睾丸 ,采用酶次第消化 ,培养过程中纯化Sertoli细胞 ,并用多种方法对Sertoli细胞进行鉴定。结果　大鼠睾丸支持细胞占培养细胞总数的 90 %以上。HE染色 ,Sertoli细胞突起很多 ,核仁清晰 ,在胞质中可见吞噬物和大小不等的空泡 ;甲基绿 派洛宁染色 ,Sertoli细胞富含RNA ;Feulgen染色和透射电镜 ,核仁周围可见卫星核小体。结论　本实验培养方法可获得更多、纯度高的Sertoli细胞 ,HE染色、甲基绿 派洛宁染色、Feulgen染色和透射电镜是鉴定Sertoli细胞的有效方法

Identification and Isolation and Purification of Testis Sertoli Cells in Rat

Objective To simplify the method of purification and culture of the sertoli cells of rat testis and to observe the sertoli cells with different methods. Methods Testis are isolated from 16～22-day old wistar rats and are subjected to sequential enzymatic treatment. The sertoli cells were purified and identified with several methods.Results More than 90% of the sertoli cells were isolated and cultured. Nucleolus are clear, grains and vacuoles can be observed in the cytoplasm with HE staining; DNA was dyed into green, RNA was dyed into red with MG-P staining. The satellite nuclear bodys were observed in the nucleus with Feulgen staining and transmission electron microscope (TEM).Conclusion More sertoli cells were obtained by this experiment method with high purification. HE, MG-P, Feulgen staining and TEM are useful methods for identiflication of the sertoli cells.