人脐带华通胶间充质干细胞的体外分离培养及鉴定

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24～48 h内细胞开始贴壁生长,5～7 d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.