摘要
目的:通过表皮干细胞对基底膜的黏附性,利用层粘连蛋白和Ⅳ型胶原作为递质,探索提取和培养表皮干细胞的技术方法。方法:取新生1～3d的Wistar大鼠全层皮肤,通过酶消化法获得幼鼠表皮,并制成单表皮细胞悬液。以层粘连蛋白和Ⅳ型胶原作为基底膜的替代物,利用表皮干细胞对基底膜的吸附性筛选表皮干细胞。结果:与角质细胞对照,表皮干细胞对层粘连蛋白和Ⅳ型胶原的吸附性很好,克隆形成率较高。用SABC法对其进行整合素β1单抗的免疫组化染色;广谱角蛋白单抗的免疫组化染色观察细胞克隆的细胞浆内出现棕黄色的阳性表达。结论:用层粘连蛋白和Ⅳ型胶原快速黏附法可从大鼠皮肤分离和富集表皮干细胞。
AIM:To investigate the extraction and culture method for epidermal stem cells( ESCs) with laminin and collagen Ⅳas transmitter through adhesiveness of ESCs on basement membrane. METHODS:The full skin of newborn Wistar rats of one to three days old was take n, and the epidermis of young rats was taken with the enzymic digestion method a nd was made into suspension.The laminin and collagen Ⅳwere taken as the substit ution of basement membrane,and the ESCs were screened by using adhesive ability of ESCs on basement membrane. RESULTS:The ESCs had better adhesive ability on laminin and collagen Ⅳ,the co lony formation efficiency(CFE) was higher than keratinocytes.The immunohistochem ical staining method was used on integrin β1 with SABC method.The brown positi ve expression was found in cytoplasm of cell colony observed by the immunohistoc hemical staining method of broad spectrum keratin. CONCLUSION:The ESCs can be separated and collected from skins of rats by using rapid adhesive method of laminin and collagen Ⅳ.
出处
《中国临床康复》
CAS
CSCD
北大核心
2005年第2期88-89,i004,共3页
Chinese Journal of Clinical Rehabilitation
基金
黑龙江省重大科技攻关项目(GAO1CO2)
黑龙江省卫生厅指导项目(2002-095)~~
