成年犬睾丸支持细胞体外分离培养的研究  

Study on the isolation and culture in vitro of Sertoli cells from adult canine testes

作　　者：葛秀国[1] 张伟[1] 丁向彬[1] 李吉霞[1] 

机构地区：[1]天津农学院动物科学与动物医学学院,天津300384

出　　处：《黑龙江畜牧兽医》2014年第8期17-20,267,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：天津农学院科技发展基金计划项目(2008D009);天津市高校优秀青年教师资助计划项目

摘　　要：为了研究犬睾丸支持细胞体外生长和生物学特征，试验采用组织培养法和酶消化法分离该细胞，并通过H．E．染色、油红O染色和AKP染色对细胞进行鉴定。结果表明：组织培养法和酶消化法均获得了成年犬睾丸支持细胞，细胞增殖活力旺盛；H．E．染色可见睾丸支持细胞呈长梭形或不规则的多边形，有3～4个突起，胞质呈浅红色；油红O染色可见支持细胞的胞质中存在脂滴，被染成枯红色，而细胞核不着色。将支持细胞与生殖细胞共培养，精原干细胞可以贴附于支持细胞表面进行体外增殖，而难以直接贴于皿底生长。AKP染色可见精原干细胞呈AKP阳性，而支持细胞呈阴性。说明成年犬支持细胞可以在体外进行培养，具有其他物种支持细胞共有的一些生物学特征。To study the growth in vitro and biological characteristics of canine Sertoli cells, these ceils were isolated using the methods ot tmsue culture and enzymatic digestion, and then were identified by hematoxylin and eosin （H. E. ）staining, oil red O staining and alkaline phospha- tase （ALP） staining. The results showed that the Sertoli cells from aduh canine testes could be obtained using both methods of tissue culture and enzymatic digestion, and the cells had strong ability of proliferation. The H. E. staining revealed that Sertoli cells were elongated spindle - shaped or irregular polygon - shaped ceils, there were three to four protrusions, and the cytoplasm was stained in light red. Oil Red O staining revealed that there were lipid droplets in the cytoplasm of the Sertoli cells, the cytoplasm was stained in organge - red, and the nucleus was not colored. When the Sertoli cells and germ cells were co - cultured, the spermatogonial stem cells could proliferate in vitro through attaching to the surfaces of the Sertoli cells, but they were difficuh to grow through directly attaching to the bottom of the dishes. AKP staining revealed that the spermatogunial stem cells were ALP - positive, and the Sertoli cells were negative. The results indicate that the adult canine Sertoli ceils can be cultured in vitro, and share some of biological characteristics of the Sertoli cells from other species.

关 键 词：犬 支持细胞 分离 体外培养 鉴定 

分 类 号：S858.292[农业科学—临床兽医学] Q813.11[农业科学—兽医学]