【摘要】 目的:用改良方法对小鼠睾丸支持细胞进行体外快速分离与培养,以期建立一种简单快捷纯度高的支持细胞原代培养方法。方法:用0.25%胰蛋白酶和0.1%胶原酶,对睾丸组织进行次第消化,用福尔根染色法和免疫细胞化学方法鉴定睾丸支持细胞。结果:细胞培养72h存活率超过90%,纯度在85%以上。福尔根染色和免疫细胞化学染色结果示,分离细胞有卫星核小体存在,且细胞中FasL阳性表达,表明分离培养的细胞确为睾丸支持细胞。结论:睾丸组织经曲细精管分离后进行胶原酶和胰蛋白酶次第消化,可快速高效分离睾丸支持细胞。更多还原

【Abstract】 Objective:To use the modified method to separate and culture sertoli cells in order to set up a simple and effective method for separating sertoli cells.Method:The tissue was treated with the 0.25% collagenase and 0.1% trypsin sequentially.Sertoli cells were identified according to Feulgen staining and immunocytochemistry.Results:The cell viability was over 90% and the purity was over 85% after 72 hours.The satellite karyosomes were stained inviolet.The expression of FasL was examined by SABC staining.Conclusion:Separating the seminiferous tubule is an efficient way for the sertoli cell digested with collagenase and trypsin sequentially.更多还原