大鼠表皮干细胞的分离培养  

Isolation and Culture of Rat Epidermal Stem Cells

作　　者：郭延敏[1,2] 杨学义[3] 段艳萍[2] 杨春荣[1] 雷安民[1] 冯秀亮[2] 

机构地区：[1]西北农林科技大学国家干细胞工程技术研究中心陕西分中心,陕西杨陵712100 [2]第四军医大学西京医院实验外科,陕西西安710032 [3]洛阳师范学院生命科学系,河南洛阳471022

出　　处：《动物医学进展》2009年第9期29-33,共5页Progress In Veterinary Medicine

基　　金：陕西省社发攻关项目(2008K14-04)

摘　　要：为探索一种简单而又高效的分离大鼠表皮干细胞（ESCs）的方法,采用酶消化法和组织块法对大鼠表皮干细胞进行分离、培养;并用Ⅳ型胶原快速黏附法筛选大鼠表皮干细胞,免疫组织化学方法检测CK15、P63、β1-integrin和α6-integrin表达情况;同时,测定ESCs单细胞克隆与克隆形成率及表皮干细胞生长曲线。结果表明,采用组织块法和酶消化法均能分离得到大鼠表皮干细胞,获得的细胞生长良好,具有较高的克隆形成率,前者可传至第6代,而后者传至3代~4代便分化;干细胞标记物CK15、P63、β2-integrin和α6-integrin均呈阳性;酶消化法和组织块法均能成功地分离、纯化和培养大鼠表皮干细胞,组织块法相对较好。The study was to explore a feasible and efficient method for isolating rat epidermal stem cells. Direct-tissue culturing and enzyme-digesting were used to isolate and culture rat epidermal stem cells. Then, the epidermal stem cells were chosen by type IV collagen. The expressions of CK15, P63, β1-integrin and α6-integrin in ESCs were detected by immunohistochemistry staining. Meanwhile, the clone forming efficiency and the growth curve of epidermal stem cells were also detected. The epidermal stem cells were got by both methods, and the epidermal stem cells had higher colony forming efficiency. The cells got by direct-tissue culturing could be passaged to the 6th generation, while the cells got by enzyme digestion were differentiated after passaged to 3-4th generation. Immuno histochemistry staining showed that CK15, P63, β1-integrin and α6-integrin were strongly expressed in the cultured epidermal stem cells. Both methods could isolate, purify and culture rat epidermal stem cells successfully, but direct-tissue culturing is more suitable for the isolation of rat epidermal stem cells.

关 键 词：表皮干细胞 分离 培养 鉴定 大鼠 

分 类 号：Q813.11[生物学—生物工程]