大鼠睾丸支持细胞的分离纯化与鉴定.pdfVIP

10 1 V ol. 10 No. 1 2004 PR OGRESS OF A NAT OM ICA L SCIENCES 2004 曹 博, 郑俊波, 郭筠秋 ( , 150086) 简化大鼠睾丸支持细胞的分离纯化方法, 通过不同方法对Sertol 细胞进行形态学观察鉴 定 取鼠 16~ 22 天的雄性W star 大鼠睾丸, 采用酶次第消化, 培养过程中纯化Sertol 细胞, 并用多 种方法对Sertol 细胞进行鉴定 大鼠睾丸支持细胞占培养细胞总数的90 % 以上HE 染色, Sertol 细 胞突起很多, 核仁清晰, 在胞质中可见吞噬物和大小不等的空泡; 甲基绿派洛宁染色, Sertol 细胞富含RNA ; Feulgen 染色和透射电镜, 核仁周围可见卫星核小体 本实验培养方法可获得更多纯度高的Sertol 细 胞, HE 染色甲基绿派洛宁染色Feulgen 染色和透射电镜是鉴定Sertol 细胞的有效方法 大鼠; 睾丸支持细胞; 分离; 纯化; 鉴定 R335+ 5 A 1006- 2947 ( 2004) 01- 00 34 - 0 3 Identification and Isolation and Purification of Testis Sertoli Cells in Rat CAO Bo, ZH ENG Junbo, GU O Yunq u ( Department of H st ology and Embryology , Harb n M ed cal U n vers t y , He longj ang Harb n 150086 Ch na) Abstract Objective T o s mpl fy the method of pur f cat on and cultur e of t he ser tol cells of rat test s and to observ e the sertol cells w th d fferent methods. Methods T est s are solated fro m 16~ 22day old w star rats and are subjected to sequent al enzymat c treatment . T he sertol cells w ere pur f ed and dent f ed w t h several met hods . Results M ore than 90 % of the sertol cells w ere solated and cultured . N ucleolus are clear, gr a ns and v acuoles can be observ ed n the cytoplasm w th HE sta n ng; DNA w as dyed nto green, R NA w as dy ed nto red w th M GP sta n ng. T he satell te nuclear bodys w er e observed n the nucleus w th Feulg en sta n ng and transm ss on electro n m croscope ( T EM ) . Conclusion M ore sertol cells w ere obta