小鼠睾丸支持细胞的原代培养和鉴定

Primary culture and identification of ICR mice sertoli cells

目的建立小鼠睾丸支持细胞培养方法,为从细胞和分子水平研究环境化合物对雄性生殖系统的影响提供条件。方法采用两步胶原酶消化,提取出生10 d的ICR雄性小鼠睾丸支持细胞,用Tris-HCl处理后除去生精细胞,进一步纯化。倒置相差显微镜和苏木精-伊红(HE)染色观察细胞生长和形态。Feulgen染色法鉴定支持细胞。结果培养的支持细胞增生活跃,纯度>95%,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。为今后研究生殖毒理提供了条件和方法。

Objective To establish culture method of ICR mice sertoli cells and to study the effects of the environmental chemicals on the male reproductive system at the cell molecular level. Methods The Sertoli cells were derived from the ten days old mice testes by twice collagenase digestion, then they were treated with Tris - HC1 to remove spermatogenic cells. To Identify the Sertoli cells with Feulgen staining, and to view the morph of cultured Sertoli cells under the microscope. Results Sertoli cells grew vigorously, the purity of the cells were over 95 percent, and bipolar Corpuscula in nucleus was clearly observed after Feulgen staining. Conclusion Twice collagenase digestion to separate Sertoli cells and Feulgen staining to identify is simple and feasible and the purity is high. The results provided a theoretical foundation for study on the male reproductive toxicity of the environmental chemicals in vitro.