小鼠睾丸支持细胞的体外分离和纯化

Separation and Purification of Testis Sertoli Cells in Mouse

目的寻求简便经济的方法分离纯化小鼠睾丸支持细胞,提高其获取量。方法用0.1%的胶原酶和0.25%的胰蛋白酶次第消化准备7-8天小鼠生殖细胞悬液,置于37℃,5%CO2孵箱内培养,4小时后换液,24小时后向培养板内加入20 mmol Tris-HCl低渗处理3分钟,去除精原细胞,即得到高纯度的支持细胞。结果在小鼠睾丸支持细胞的分离培养中,支持细胞占培养细胞90%以上。结论应用本实验方法分离小鼠睾丸支持细胞获得成功并简化了国内现行的分离方法。

Objective To search for a simple and economical method to separate and purify mouse testis sertoli cells so as to increase the quantity of sertoli cells. Methods 0. 1% collagenase and 0. 25 % typsin was sequentially used to prepare germ cell suspension of mouse at 7--8days. Cell suspensions were incubated at 37℃ in 5% CO2. After 4 hours replenished the culture medium and after 24 hours added 20 mmol Tris-HCl into culture wells for 3 minutes, almost all the spermatogonia were removed and high purity sertoli cells were obtained. Results The isolation and culture cells contained greater than 90% sertoli cells. Conclusion The application of this method used in the present experiment has been successful in getting mouse testis sertoli cells and simplified domestic current methods.