人脐血MSCs的分离培养及向神经的诱导分化  

Inducing mesenchymal stem cells isolated from human umbilical cord blood into neuron-like and glia-like cells

作　　者：曲静[1] 张稷[1] 於葛华[1] 孙万平[1] 张洪涛[2] 惠国桢[3] 张学光[1] 

机构地区：[1]江苏省干细胞研究重点实验室 [2]苏州大学附属第一医院骨科 [3]苏州大学附属第一医院神经外科

出　　处：《江苏医药》2006年第10期909-911,共3页Jiangsu Medical Journal

基　　金：国家自然科学基金(30271325);江苏省自然科学基金(BK2001170)

摘　　要：目的探讨人脐血中分离所得到间充质干细胞（MSCs）向神经组织细胞诱导分化的实验条件。方法淋巴细胞分离液分离正常人脐血单个核细胞，利用差速消化法通过多次传代得到MSCs，流式细胞仪测定细胞表型。取2至4代的MSCs加入含有维甲酸（RA）与神经生长因子（NGF）的诱导分化培养基将其定向诱导，免疫细胞化学染色检测诱导后的MSCs神经元和神经胶质细胞标志的表达。结果脐血MSCs每传一代，细胞数量增加约2～3倍。流式细胞仪检测，脐血MSCs不表达CD34，表达CD25、CD29、CD105、CD106。诱导后的细胞不仅形态类似神经元和神经胶质细胞，而且表达神经丝蛋白200（NF）、神经元特异性烯醇化酶（NSE）和胶质原纤维酸性蛋白（GFAP）。结论从人脐血中可以分离得到与骨髓相似的MSCs，并且能够在体外分化为神经元样和神经胶质细胞样细胞。Objective To isolate and induce mesenchymal stem cells（MSCs） from human umbilical cord blood into neuron-like cells in vitro. Methods Human cord blood samples were obtained sterilely. The MSCs were isolated by lymphocyte separation medium, and purified by the different sensitivity to trypsine. The phenotype of MSCs was analyzed by flow cytometry. The 2nd to the 4th passage of MSCs was induced into neuron-like and gila-like cells with ＂differentiated medium＂ consisting of all-trans-retinoic acid（RA） and human nerve growth factor（NGF） in vitro. Specific marker was detected by immunocytochemistry method. Results The number of MSCs was increased by two-to three times with each expanded passage. Flow cytometry analysis showed that MSCs did not express CD34, but expressed CD25 ,CD29 and CD105 consistedly with MSCs from bone marrow. The induced MSCs were neuron and glia-like morphologically and expressed neurofilament （NF）, neuron-specific enolase（NSE）, and glial fibrillary acidic protein（GFAP） verified by imrnunocytochemistry method. Conclusion MSCs can be isolated from human umbilical cord blood and differentiated into neuron-like and gila-like cells.

关 键 词：人脐血 间充质干细胞 诱导分化 神经元样细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]