大鼠睾丸支持细胞的分离培养及鉴定  

Isolation culture and identification of testis sertoli cells from rat

作　　者：丁天云[1] 张明[2] 杨洋[1] 简少卿[1] 阳刚[1] 胡宝庆[1] 文春根[1] 

机构地区：[1]南昌大学生命科学学院,江西南昌330031 [2]江西生物科技职业学院,江西南昌330200

出　　处：《南昌大学学报（理科版）》2016年第5期493-496,510,共5页Journal of Nanchang University(Natural Science)

基　　金：国家自然基金资助项目(21467015);南昌大学研究生创新专项资金(cx2015098)

摘　　要：通过简化、优化现有的SD大鼠睾丸支持细胞分离纯化方法,获得高纯度大量的支持细胞。选用15d的雄性SD大鼠,处死取出睾丸,采用0.25%胰蛋白酶消化的一步消化法分离支持细胞,置于35℃、5%CO2和100%湿度空气的CO2培养箱中进行培养。24h后用20mmol·L-1的Tris-HCl处理细胞,并用油红O、HE染色和Feulgen染色观察对培养的支持细胞进行鉴定。分离纯化的支持细胞纯度达到90%以上,纯化获得的细胞其形体结构与支持细胞一致。因此,一步消化法可以获得高纯度的支持细胞。The existing methods of isolation and purification was simplified and optimized for testis sertoli cells of SD rat,and the high purity sertoli cells was obtained. Male SD rats on 15 d were utilized and execu- ted to remove testicles. The sertoli cells were separated from testicles by one-step digestion method using lg/L collagenase,0.25% trypsin,which were cultured under 35 ℃ ,5% CO2 and 100% humidity air. After 24 hours,the sertoli cells were handled by 20 mmol · L-1 Tris-HCl,and were stained and identified by oil red O, HE and Feulgen staining. The purity of separated and purified the sertoli cells was more than 90%, and the morphological structure of the cells was in accordance with the sertoli cells. Therefore, the one-step digestion method could be used and got high purity sertoli cells.

关 键 词：SD大鼠 支持细胞 分离 鉴定 

分 类 号：Q813.11[生物学—生物工程]