人骨髓间充质干细胞分离培养及其向神经细胞分化的研究

目的探讨人骨髓间充质干细胞(BMMSCs)向神经细胞分化的潜能。方法体外培养人BMMSCs,并诱导其向神经细胞分化。诱导前后观察细胞形态变化,免疫细胞化学法检测细胞神经元特异性烯醇化酶(NSE)的表达以鉴定细胞类型,流式细胞术检测细胞周期以确定细胞增殖活性。结果分离培养的BMMSCs有克隆增殖能力。诱导后,分化细胞呈现典型神经元的表型,且免疫荧光染色呈NSE阳性。流式细胞术分析显示诱导前BMMSCs的增殖指数(PI)较高,细胞增殖活跃。与诱导前比较,诱导分化细胞的PI显著降低(P<0.01),细胞增殖发生抑制,而分化阶段各期细胞比例差异无统计学意义(P>0.05)。结论 BMMSCs可以在体外增殖扩增,并能分化为神经元样细胞,且分化的神经元样细胞能较长时间存活。

Isolation,cultivation and neural differentiation of human bone marrow mesenchymal stem cells

Objective To explore the neural differentiation potential of human bone marrow mesenchymal stem cells （BMMSCs）. Methods The human BMMSCs were cultured and induced to differentiate into neural cells in vitro. Before and after induction, the morphology of all tested cells in the course of cultivation were observed. The expression of neuron specific enolase（NSE） was measured to detect cell type by immunocytochemical method. And flow cytometry was used to determine the cell cycle phases and detect proliferative activity of these cells. Results BMMSCs which were isolated from the human marrow had the ability of proliferation. After induced differentiation, the differentiated cells showed typically neuronal morphology and were NSE-positive. Meanwhile, flow cytometry analysis showed high proliferative index （ PI）, which indicated that BMMSCs were actively dividing. The PI decreased significantly after fieural differentiation compared with that before induction （P 〈 0. 01 ）. However, the percentage of cells in cell cycle phases among these stages of differentiation had no statistical significance （P 〉 0.05 ）. Conclusion BMMSCs can be isolated, pibliferated and further be induced into neural-like cells, which have a potential of long-time survival in vitro.