摘要
目的:探讨睾酮对体外培养小鼠精原干细胞增殖与分化的影响。方法:采用Percoll密度梯度分离及差速贴壁法纯化精原干细胞,用免疫荧光染色及流式细胞检测对小鼠精原干细胞进行鉴定。设置实验组和对照组,两组均以支持细胞作为饲养层培养精原干细胞,在实验组DMEM/F12完全培养液中添加睾酮。用酶联仪检测细胞的生长情况;用流式细胞仪对细胞生长周期进行判断;采用ICSI技术让精子细胞与卵母细胞结合,观察卵裂细胞数,体外培养3天后进行染色体形态与数量分析。结果:在添加睾酮的培养基中培养的精原干细胞,其OD值增长较快(P<0.05);精原干细胞DNA合成期(S期)的含量增多(P<0.05);精子细胞与卵子结合以后可得到二倍体配子。结论:睾酮能促进精原干细胞的体外增殖与分化。
Objective：To investigate the effect of testosterone on proliferation and polarizationof rat spermatogonial stem cells in vitro culture. Methods：The pereoll discontinue density gradient centrifugation and differential attachment method were used to purify the spermatogonial stem cells. Rat spermatogonial stem cells was con- firmed by immunofluoreseence and flow cytometry （FCM）. There were control group and expriment group, the spermatogonial stem cells were cultured on sertoli cells. Testosterone was added to DMEM/ F12 medium. The cell growth of spermatogonial stem cells was determined by enzyme linked immunosorbenl assay （ELISA）. The cell cycle of spermatogonial stem cells was analyzed by flow cytometry; Sperm allied with ovum by intracytoplasmic sperm injection （ICSI）, and it＇s chromosome figure and quantity was examined after three days. Results：OD value of spermatogonial stem cells gradually increased in medium eontaing testosterone（P〈0.05）, and the quantity of spermatogonial stem gradually increased in DNA synthesizing stage （S stage）（P〈0.05） ; diploid gamete can be obtained after integration of sperm and ovum. Conclusions： Testosterone can improve the proliferation and polarization of the spermatogonial stem cells in vitro.
出处
《临床泌尿外科杂志》
北大核心
2009年第6期460-462,467,共4页
Journal of Clinical Urology
基金
贵州省社会发展攻关基金资助项目(No:黔科合S子[2007]1045)
关键词
精原干细胞
睾酮
体外培养
小鼠
spermatogonial stem cells
testosterone
vitro culture
rat
