【摘要】 目的探讨在体外诱导成人骨髓间充质干细胞分化为神经元样细胞的方法。方法采用梯度离心法和全骨髓法,利用含血清的DMEM培养基培养成人骨髓间充质干细胞,以β-巯基乙醇诱导其向神经元样细胞方向分化。应用免疫细胞化学技术对培养细胞及其分化细胞进行鉴定。结果骨髓细胞培养至第10d时,全骨髓法收获的细胞数为(1.73±0.44)×107/ml,而梯度离心法收获的细胞数为(7.65±0.52)×107/ml,其差异有统计学意义(P<0.01);但两种方法所得细胞的生物学特性没有差别。骨髓间充质干细胞经β-巯基乙醇诱导后细胞形态发生改变,伸出突起并交织成网;免疫细胞化学法检测显示54.76%±3.65%的细胞表达神经元特异性蛋白NSE,同时36.28%±4.27%的细胞表达神经胶质细胞特异性蛋白GFAP。结论梯度离心法较全骨髓法收获细胞数量大。在体外条件下,利用β-巯基乙醇和适宜的培养液可使骨髓间充质干细胞定向分化为神经元。更多还原

【Abstract】 Objective To explore a way that induce human bone marrow mesenchymal stem cells to differentiate into neuron-like cells in vitro.Methods Bone marrow mesenchymal stem cells were cultivated in a medium containing serum by using density gradient centrifugation method and whole marrow method and were induced to differentiation by βmercaptoethanol.Immunohistochemistry were used to identify the type of cells.Results The number of cells was（（1.73）±（0.44））×10⁷/ml by using whole marrow method and （（7.65）±（0.52））×10⁷/ml by using density gradient centrifugation method at the 10th day cultivation respectively.There was statistically significant difference（P<（0.01））.After the induction,bone marrow mesenchymal stem cells displayed processes that formed extensive networks.Positive cells for NSE were（54.76%）±（3.65%）,and for GFAP were（36.28%）±（4.27%） respectively.Conclusions The number of cells was more with density gradient centrifugation method than with whole marrow method.Bone marrow mesenchymal stem cells could eventually differentiate into neuronlike cells in vitro by β-mercaptoethanol and optimal medium.更多还原