人表皮干细胞的分离、纯化培养及鉴定  

Isolation,culture and identification of human keratinocyte stem cells

作　　者：马英智[1,2] 王心蕊[3] 何旭[1] 牛云[4] 李玉林[1] 

机构地区：[1]吉林大学白求恩医学院病理生物学教育部重点实验室,吉林长春130021 [2]空军航空大学飞行基础训练基地门诊部 [3]吉林大学人兽共患病研究所 [4]中日友好医院病理科

出　　处：《中国实验诊断学》2009年第2期143-145,共3页Chinese Journal of Laboratory Diagnosis

基　　金：国家863重大专项(2004AA205020);教育部博士学科点专项科研基金(20020183064);国家自然科学基金(30700872)

摘　　要：目的对人表皮干细胞(human keratinocyte stem cells,hKSCs)进行分离、纯化培养及鉴定,探索一种高效分离及纯化培养hKSCs的途径,为组织工程皮肤的构建提供种子细胞。方法利用酶消化法分离hKSCs,快速粘附法纯化hKSCs并进行无血清培养,倒置相差显微镜观察形态变化;以角蛋白19(cytokeratin,CK19)及整合素β1、p63为一抗,利用激光扫描共聚焦显微镜(laser scanning confocal microscope,LSCM)对原代培养的细胞进行免疫细胞化学检测。结果经快速粘附法筛选的细胞在相差显微镜下观察形态一致,呈上皮样生长;LSCM下CK19及整合素β1、p63均呈阳性反应,初步证实分离出的细胞为hKSCs。结论成功建立了hKSCs分离、纯化和培养的方法,为组织工程皮肤的研究提供了理论与实验基础。Objective To explore a new method of the isolation, culture of human keratinocyte stem cell （hKSCs） in vitro for the provision of scedceUs in tissue engineering of skin. Methods Epidermis was dissociated into single cells with trypsin and dispase. The rapidly adherent cells were cultttred by DK-SFM and observed under phase contrast microscope,and they were identified with immunocytcehemistry by laser scanning confceal microscope （LSCM）. Results With phase contrast microscope, the rapidly adherent cells were observed to uniformed population of cells and epithelial-like. LSCM showed the rapidly adherent cells were positive for βlintergrin, cytokerafin 19 and 1363. Conclusion This study shows that human keratinocyte stem cells could be isolated and cultured in vitro successfully.

关 键 词：表皮干细胞 分离 培养 鉴定 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]