人脐带间充质干细胞的分离培养及生物学特性  

Isolation and cultivation of MSCs from human umbilical cord and identification of their biological characteristics

作　　者：孙丽[1] 于丽[1] 张华芳[2] 江洪[2] 

机构地区：[1]潍坊医学院组织学与胚胎学教研室,潍坊261053 [2]潍坊市人民医院保健三科,潍坊261041

出　　处：《解剖科学进展》2011年第2期131-134,共4页Progress of Anatomical Sciences

基　　金：山东省自然科学基金项目(NO.Y2008C26);山东省教育厅基金资助项目(NO.J07YD01);潍坊医学院研究生创新基金项目(NO.YC2008011)

摘　　要：目的建立一种稳定、高效的分离培养人脐带间充质干细胞(MSCs)的方法,并对其生物学特性进行鉴定。方法分别采用组织块贴壁法和双酶消化法分离培养人脐带MSCs,对比其培养成功率;流式细胞术检测脐带MSCs表面抗原及细胞周期;采用丹参联合生长因子的方法诱导其向神经细胞分化,观察脐带MSCs向神经元分化潜能。结果组织块贴壁法获得脐带MSCs成功率高;细胞表达CD29、CD44和CD90,不表达造血干细胞表型CD34。细胞倍增时间为30h,G0-GI期和S+G2+M期所占比例分别为78.47%和21.53%。脐带MSCs在体外经诱导分化出现神经元样细胞改变,且表达神经元特异性烯醇化酶(NSE)。结论成功建立了高效稳定的脐带MSCs分离培养方法,脐带MSCs具备较强的分化潜能和增殖能力,为临床移植治疗提供了理想的细胞来源。Objective To establish a stable and efficient method for separating mesenchymal stem cells（MSCs） from human umbilica1cord,and identify their biologica1 characteristics. Methods MSCs were isolated from human umbilica1 cord with tissue adherent method and double enzyme digestion method respectively. Cell cycles and surface antigens were tested by flow cytometry. The third passage MSCs were induced into neuron-like cells by Danshen and growth factor and characterized by neuron specific enolase（NSE） immunohistochemistry. Results More percentage of MSCs were obtained by tissue adherent method than by double digests method,flow cytometry analysis showed that MSCs expressed CD29-,CD44-and CD90-positively,but negatively for CD34. Cel1 cycle analysis showed that the proportions of G0-GI phase and S ＋ G2 ＋ M phase were 78.47% and 21.53% respectively. MSCs could be induced into neuron-like cells and showed NSE positive. Conclusion MSCs derived from human umbilica1 cord have differentiated and proliferative capacity,which could be used for clinical cell transplantation.

关 键 词：脐带 间充质干细胞 培养 生物学特性 人 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]