小鼠表皮干细胞的分离与培养  

Isolation and Cultivation of Mouse Epidermal Stem Cells

作　　者：王丽娟[1] 孙耀兰[1] 杨雪[1] 王友亮[1] 杨晓[1] 

机构地区：[1]军事医学科学院生物工程研究所蛋白质组学国家重点实验室,北京100071

出　　处：《生物技术通讯》2011年第3期389-393,共5页Letters in Biotechnology

基　　金：国家重大新药创制项目(2009ZX09501-027)

摘　　要：目的:探讨体外分离和培养小鼠表皮干细胞和分析表皮干细胞克隆形成能力的方法。方法:采用中性蛋白酶和胰酶消化新生小鼠表皮基底层细胞,将细胞直接接种在细胞瓶中,在无滋养层条件下培育;利用表皮干细胞标记物K15和α6整联蛋白进行免疫荧光鉴定;以小鼠胚胎成纤维细胞作为滋养层与成年小鼠角质细胞共培养,进而分析表皮干细胞的克隆形成能力。结果:新生小鼠表皮干细胞克隆在培养2～3 d后开始形成,细胞核质较小,细胞呈小而圆的形态特征;传代后的细胞可以被K15和α6整联蛋白特异性标记。结论:利用该方法能够实现对小鼠表皮干细胞的体外培养和传代。Objective：To isolate and culture the mouse epidermal stem cells and analyze its capability of clone formation for further studies.Methods：The neonate mice epidermal basal cells were digested by dispase and trypsin and then seeded directly and cultivated in culture flasks without any feeder cells.The epidermal stem cells were identified by K15 and α6-integrin staining and their clone formation abilities were evaluated when they were co-cultured with mouse embryonic fibrolasts feeder cells in a cell differentiation condition.Results：The clones of neonate mice epidermal stem cells were successfully formated after 2～3 day＇s cultivation characterized by low nucleo-cytoplasma ratio and tiny and round shape.Those cells could be specialized marked with K15 and α6-integrin after cell passage.Conclusion：The cultivation and passage of mouse epidermal stem cells can be achieved using this kind of method.

关 键 词：小鼠表皮干细胞 培养 传代 克隆形成 

分 类 号：Q813.11[生物学—生物工程]