大鼠骨髓间充质干细胞体外分化为神经元样细胞的研究

目的：研究大鼠骨髓间充质干细胞(rMSCs)体外扩增不同诱导剂向神经元样细胞诱导分化的潜能。方法：采用贴壁筛选法分离Wistar大鼠股骨rMSCs，体外扩增5-10代后，分别用2-巯基乙醇、硫代甘油等无血清的DMEM培养液诱导hMSCs分化为神经元。用流式细胞仪鉴定rMSCs特异抗原表达，免疫组化鉴定诱导后神经元样细胞的神经元烯醇化酶(NSH)、神经丝蛋白(NF)、胶质纤维酸性蛋白(GFAP)。结果：大鼠MSCs体外扩增传至5代后，流式细胞仪显示rMSCsCD27、CD44、CD90呈阳性表达；将rMSC加入2-巯基乙醇和硫代甘油等诱导剂3h后，rMSC胞体收缩，突起伸出；免疫组化显示诱导出的神经元样细胞NSE、NF表达阳性，GFAP阴性。结论：大鼠骨髓间充质干细胞在体外-定条件下可以分化为神经元样细胞。

In vitro study on neuron-like cells differentiated from adult Wistar rat mesenchymal stem cells

Objective:To investigate neuron-like cells dif fe rentiated from Wistar rat mesenchy-mal stem cells(rMSCs).Methods:rMSCs wer e obtained from femurs marrow and multiblicated in culture medium.The surface a ntigen expression of rMSCs was detected by flow cytometry.rMSCs were induced to differentiate into neuron-like cells with DMEM/basic fibroblast growth fact or and2-mercaptoethanol or DMEM/monothioglycero1,respectively.Specific mar kers and stuctures were detected by immunohis-tochemistry and histochemistry.N euron-specific enolase(NSE),nestin,neurofilament(NF),glial fibrillary ac idic protein(GFAP)were detected by immunohistochemistry.Results:The isolated and cultured rMSCs comprised single phenotypic population and displayed a fibr oblast-like morphology.rMSCs were multi-plicated to be undifferentiated cell s in culture for more than5passages.Flow cytometry showed that98.9%,98 .5%,99.8%rMSCs expressed strongly antigens CD27,CD44,CD90which was identical to rat bone marrow-derived rMSCs.Most cells exhibited typical n euron-like phenotye after induction.Immuno-histochemistry staining showed tha t the induced different-passage MSCs expressed neuron-specific eno-lase(NSE ),nestin,neurofilament(NF),but did not express glial fibrillary acidic pr otein(GFAP).Con-clusion:rMSCs can be differentiated into neurons in vitro .