人脐带华通胶间充质干细胞的体外分离培养及鉴定  

Isolation,Characterization and Induced Differentiation of Human Umbilical Cord Wharton's Jelly-derived Mesenchymal Stem Cells

作　　者：胡晶琼[1] 李慧玉[1] 欧阳为相[2] 胡丽[3] 卢聪[1] 张兰男[1] 陈莉莉[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院干细胞中心,武汉430022 [2]华中科技大学同济医学院附属协和医院妇产科,武汉430022 [3]华中科技大学同济医学院附属协和医院口腔医疗中心,武汉430022

出　　处：《华中科技大学学报（医学版）》2012年第5期529-533,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金国际合作与交流项目(No.31110103905);国家留学回国人员启动基金项目

摘　　要：目的研究由脐带华通胶组织(Wharton’s jelly)在体外分离、培养、扩增获得脐带间充质干细胞(umbilical cord derived mesenchymal stem cells,以下简称UC-MSCs)的方法,并进行UC-MSCs的各项鉴定。方法脐带华通胶组织采用胶原酶以及胰酶序贯消化法分离得到UC-MSCs,用流式细胞仪分析其表型特征,向成骨、成脂以及成神经元细胞诱导分化并鉴定。结果由人脐带华通胶可以有效获得间充质干细胞。原代细胞培养24～48h内细胞开始贴壁生长,5～7d左右可以传代培养,在2～3周时间内可以迅速扩增至107到108数量级。各项分化鉴定试验证实UC-MSCs具有多向分化潜能,能分化成骨、成脂细胞以及神经元细胞。UC-MSCs在体外培养传至20～30代仍保持稳定的细胞表面标记。结论由人脐带华通胶可以有效地获得间充质干细胞,这种UC-MSCs能在体外长期传代培养,生物学特性稳定,具有多向分化的潜能,是今后细胞治疗很有前景的种子细胞。Objective To isolate and characterize mesenchymal stem cells derived from human umbilical cord Wharton＇s jelly（UC-MSCs）.Methods MSCs were derived from human umbilical cord Wharton＇s jelly using a consecutive enzyme digestion method of both collagenase and trypsin.FACS analysis was done to access the phenotype of UC-MSCs.Moreover,the UC-MSCs were induced to differentiate into chondrocytes,adipocytes and neurons.Results MSCs could be efficiently derived from human umbilical cord Wharton＇s jelly.Primary cells could attach around 24—48 h after plating.Around 5—7 days the first passage could be made.Primary cells could be expanded within two-three weeks in 107 to 108 range.Human umbilical cord Wharton＇s jelly MSCs could be efficiently induced to differentiate into chondrocytes,adipocytes and neurons.Phenotype of UC-MSCs was stable even after prolonged ex vivo cell culture of 20—30 passages.Conclusion We described the isolation,maintenance,ex vivo expansion and characterization of MSCs derived from human umbilical cord Wharton＇s jelly.These MSCs are multi-potent,extremely stable in ex vivo culture,and can be efficiently induced to a neuronal phenotype.These UC-MSCs hold great promise for future cell based therapy.

关 键 词：间充质干细胞 脐带 华通胶 诱导分化 

分 类 号：R394.26[医药卫生—医学遗传学]