大鼠睾丸间质细胞的原代培养、鉴定与功能监测  

Primary Culture,Identification and Functional Study of Rat Leydig Cells

作　　者：应锋[1] 龚昳[1] 孙佳音[1] 沈洁[1] 韩晓冬[1] 

机构地区：[1]南京大学医学院免疫与生殖生物学实验室,江苏南京210093

出　　处：《中华男科学杂志》2008年第1期7-10,共4页National Journal of Andrology

基　　金：国家自然科学基金(20577019)

摘　　要：目的:采用改良方法对大鼠睾丸间质细胞进行分离、纯化和原代培养,以得到更高纯度和稳定的间质细胞原代培养体系。方法:采用酶消化法分离大鼠睾丸间质细胞,再用Percoll连续密度梯度离心除去生精细胞、支持细胞等杂细胞,实现进一步纯化;用3β-HSD特异性染色法鉴定间质细胞,同时采用放射免疫法测定间质细胞睾酮分泌活性。结果:培养的间质细胞纯度达到95%以上,每个睾丸可获取间质细胞约1×106个;通过3β-HSD特异性染色鉴定,该间质细胞胞质呈蓝黑色,而且细胞具有分泌睾酮的活性。结论:酶消化后以Percoll连续密度梯度离心可分离得到高纯度且具有睾酮分泌活性的间质细胞,操作简单易行,实验方法稳定。Objective： To set up a stable primary culture system of Leydig cells with higher purity. Methods： We separated Leydig cells from other testicular cells, such as Sertoli and germ cells, by enzymatic digestion in combination with Percoll density gradient centrifugation and identified Leydig cells by 3β-HSD staining. Results： The purity achieved by this method was above 95% and the total number of Leydig cells obtained from one testicle was about 1×10^6. The cytoplasm of Leydig cells was stained in deep blue by 3β-HSD staining, and these cells possessed testosterone-secreting capability. Conclusion： Leydig cells can be separated by enzymatic digestion combined with Percoll density gradient centrifugation, and 3β-HSD staining to identify Leydig cells is simple and feasible with high purity.

关 键 词：间质细胞 Percoll密度梯度离心 原代培养 3β-HSD染色 大鼠 

分 类 号：Q813[生物学—生物工程]