成人骨髓间充质干细胞的培养及其向神经元样细胞诱导分化的实验研究  

Cultivation of human bone marrow mesenchymal stem cells and induced differentiation into neuron-like cells

作　　者：陈刚[1] 秦尚振[1] 马廉亭[1] 雷霆[2] 潘力[1] 薛德麟[2] 

机构地区：[1]广州军区武汉总医院神经外科,430070 [2]华中科技大学同济医学院附属同济医院神经外科

出　　处：《中华器官移植杂志》2006年第2期107-110,共4页Chinese Journal of Organ Transplantation

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。Objective To explore a way that induce cells to differentiate into neuron-like cells in vitro. Methods human bone marrow mesenchymal stem 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 β-mercaptoethanol. Immunohistochemistry were used to identify the type of cells. Results The number of cells was （1.73 ± 0. 44）×10^7/ml by using whole marrow method and （7. 65 ± 0. 52） ×10^7/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. 280/00 ± 4. 270/40 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 neuron-like cells in vitro by β-mercaptoethanol and optimal medium.

关 键 词：骨髓 间质干细胞 细胞培养技术 神经元 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]