体外诱导骨髓基质细胞向神经细胞分化的形态学观察

目的:探讨骨髓源性神经干细胞分化为神经细胞的可行性,为中枢神经组织损伤修复寻找种子细胞的理想来源.方法:无菌取成大鼠长骨骨髓,密度梯度离心分离后获得骨髓基质细胞(BMSC),在神经干细胞培养液及细胞因子等条件下诱导其分化为神经元样细胞.结果:增殖培养24～72h见细胞分裂像和克隆单位;继续增殖培养仍可见细胞克隆(干细胞特性);诱导培养第10日有成熟神经细胞形成.说明BMSC在体外培养条件下,经过多种因子的"程序性"作用,可以向神经干细胞、成熟神经元及胶质细胞方向分化.结论:在适宜培养条件及诱导因子联合作用下,BMSC可成功诱导出神经元样细胞.

Morphological study on in vitro inducement and differentiation of nerve cells derived from rat BMSC

AIM: To explore the possibilities of differentiating bone marrow stromal cells (BMSC) into nerve cells (NC) and to establish the method and condition of BMSC culture in vitro, so as to use the NC derived from BMSC as germ cells in tissue-engineering. METHODS: Under sterile conditions, bone marrow stem cells, from bone marrow of SD rat and separated by gradient density centrifugation, were cultured, induced and differentiated into NSC and neuronal-like cells with cytokines consisting of retinoic acid, glial-derived neurotrophic factor and special medium confected in our laboratory (patent number: 02134314.4). RESULTS: When BMSC were proliferatively cultured for 24-72 h, cleavage phase and cluster-like clones were seen. When they were cultured in the proliferation medium, the cells could be cloned, which was one of characters of stem cells. On the 10th d after adding inducing factors into the culture medium, the nerve cells or neuron-like cells appeared. CONCLUSION: BMSC can differentiate into some cells with characteristics of nervous cells.