摘要
目的 简化大鼠睾丸支持细胞的分离培养方法 ,提高支持细胞的获取量。方法 取鼠龄 16～ 2 2天的Wistar大鼠 ,去除睾丸被膜 ,剪碎后用 0 .2 5 %胰蛋白酶和 0 .1%胶原酶次第消化。然后将其制成细胞悬液并分装入 2 5cm2 的培养瓶中 ,放置在 39℃ 5 %CO2 的培养箱孵育 48h ,以利于去除精原细胞。取出后冲洗两次 ,换液并移入 37℃ 5 %CO2培养箱进行培养。在此期间每天用倒置显微镜进行观察 ,并于培养第 6天做光镜和电镜观察。结果 在大鼠睾丸支持细胞的分离培养中 ,支持细胞占培养细胞总数的 90 %以上 ,大大提高了支持细胞的获取量。结论 应用本实验方法获取大鼠睾丸支持细胞获得成功并简化了国内现行的方法 。
Objective To simplify the method of isolation and culture of rat testis sertoli cells and increase the quantity of sertoli cells.Methods Decapsulated rat testis from 16～22 day old Wistar rats were subjected to sequential enzymatic treatment using 0.25% trypsin and 0.1%collagenase.The resulting tissues were made into cell suspantions and distributed into 25cm 2 flasks.Cell cultures were incubated at 39℃ in 5% CO 2 95% air for 48h to facilitate the removal of germ cells.After two washing and replenishing culture medium,cell culture wells were returned to the CO 2 injected incubator at 37℃ in 5% CO 2.During the couse,observed the culture cells with phase contrast microscopy daily and identified their morphology with light microscopy and electron microscopy on the sixth culture day.Results The isolation and culture cells contained greater than 90% sertoli cells and increased the quantity largely.Conclusion The application of this method used in the present experiment has been successful in getting rat testis sertoli cells and simplified domestic current methods.
出处
《哈尔滨医科大学学报》
CAS
2001年第3期185-187,F003,共4页
Journal of Harbin Medical University
