成体表皮干细胞的分离培养与鉴定  

Isolation,culture and identification of adult epidermal stem cell

作　　者：丁伟[1] 王云龙[1] 李敏[1] 端龙胜[1] 王剑[1] 

机构地区：[1]皖南医学院附属弋矶山医院烧伤科,安徽芜湖241001

出　　处：《皖南医学院学报》2010年第4期252-254,共3页Journal of Wannan Medical College

摘　　要：目的:探讨人表皮干细胞的体外快速分离培养及鉴定方法。方法:运用DispaseⅡ酶和胰蛋白酶两步法从手术切除的人包皮组织中分离获得表皮干细胞。倒置显微镜下观察培养细胞的生长状况;检测细胞克隆形成率及克隆维持时间;免疫组化染色观察表皮干细胞标志物β1整合素和角蛋白19(K19)的表达。以角质形成细胞作为对照。结果:组织学观察显示,培养24h后细胞呈克隆状生长;所分离、培养细胞的克隆形成率高于对照角质形成细胞组;免疫组化染色显示,培养细胞β1整合素及K19均呈阳性表达。结论:成体表皮干细胞在体外得到成功分离与培养。Objective：To explore a method for quickly isolating,culturing and identifying epidermal stem cells of human. Methods：Epidermis was obtained by digesting human foreskin with DispaseⅡand Trypsin-EDTA,with observation of the cell growth by inverted microscope and examination of cell cloning efficiency and duration of clone sustaining. Immunocytochemistry was used to observe the expression of β1-integrin and keratin19（K19）. Keratinocytes were served as controls. Results：Histological findings showed that colonies were formed 24 hours after inoculation. The efficiency of isolated and cultured cell cloning was higher than that of the control group. Positive expression was detected in β1-integrin and K19 of cultured cells by immunocytochemistry. Conclusion：Adult epidermal stem cells were successfully isolated and cultured in vitro.

关 键 词：表皮干细胞 细胞培养 细胞分离 鉴定 

分 类 号：Q813.11[生物学—生物工程]