新生犊牛睾丸支持细胞的高效分离培养与鉴定

Efficient separation,culture and identification of Sertoli cells in new-born calf

【目的】采用改良方法对犊牛睾丸支持细胞进行体外快速分离培养,以期建立一种简单快捷的支持细胞原代培养方法。【方法】采用1.0 g/L的Ⅳ型胶原酶和2.5 g/L的胰蛋白酶,对经分离曲细精管和组织剪碎2种方法处理的睾丸组织进行次第消化,比较2种方法对睾丸支持细胞分离效果的影响;采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】睾丸组织采用分离曲细精管后进行酶消化的时间(8 min)较组织剪碎法(15min)短,并且前者消化所得有效细胞数(1.0×107)高于后者(0.81×107),两者具有显著差异(P<0.05);福尔根染色和免疫细胞化学染色结果显示,分离细胞有卫星核小体存在,且细胞中ABP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,有助于快速高效分离睾丸支持细胞。

【Objective】 We used 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 methods of separating seminiferous tubule and cutting tissue respectively.Then,the disuse was digested with the 1.0 g/L collagenase and 2.5 g/L trypsin sequentially.The effect of the two methods on the Sertoli cell separation was compared.Sertoli cells were identified according to Feulgen staining and immunocytochemis...