音猬因子体外诱导人骨髓基质干细胞转化为神经元样细胞的研究

目的：研究体外使用音猬因子（sonic hedgehog，SHH）诱导人骨髓基质干细胞（BMSCs）分化为神经元样细胞（神经干细胞、神经细胞、神经胶质细胞）的可行性。方法：从健康志愿者髂骨抽取骨髓5ml．离心、分离BMSCs。接种于含10％胎牛血清（FBS）的DMEM／F12培养液中，BMSCs体外扩增、纯化到第五代后分别种于含有不同浓度诱导液的24孔板中：A组为空白对照；B组加SHH 700ng/ml，碱性成纤维生长因子8（fibroblast growth factor-8，FGF-8）140ng／ml；C组加SHH500ng/ml，FGF-8100ng／ml；D组加SHH250ng/ml，FGF-8 50ng／ml；E组加SHH125ng／ml，FGF-825ng／ml，诱导BMSCs分化。使用免疫组化及免疫荧光鉴定微管相关蛋白（MAP-2）、胶质纤维酸性蛋白（GFAP）、神经元烯醇化酶（NSE）的表达。结果：诱导7d后，部分BMSCs胞体收缩、突起伸出，表现出神经元样细胞的形态。除A组外其余各组免疫组化及免疫荧光染色NSE、MAP-2均为阳性，各组免疫组化及免疫荧光染色均有GFAP阳性细胞存在，且以B组、C组的MAP-2、GFAP、NSE染色阳性细胞数最多。结论：人BMSCs具有较强的自我更新和多向分化潜能，一定浓度的SHH可以在体外有效诱导人BMSCs转化为神经元样细胞。

Differentiation of bone marrow stromal cells into neuron-like cells induced by sonic hedgehog

Objective:To investigate the possibility of inducing human bone marrow stromal cells(BMSCs) into the neural-like and/or glia-like cells by sonic hedgehog(SHH).Method:BMSCs harvested from normal human marrow were passaged and purifed in DMEM/F12 media containing 10% FBS.After five passage,the purifed human BMSCs were induced by SHH with different concentrations(group A,control normal group;group B,SHH 700ng/ml,FGF-8 140ng/ml;group C,SHH 500ng/ml,FGF-8 100ng/ml;group D,SHH 250ng/ml,FGF-8 50ng/ml;group E,SHH 125ng/ml,FGF-8 25ng/ml) and detected by immunocytochemistry staining for GFAP?MAP-2?NSE.Result:After the induction of 7 days,some BMSCs cells exhibited immunoreactivity.NSE and MAP-2 positive cells exist in all groups except of control group,GFAP positive cells exist in all groups.The amount of NSE?MAP-2?GFAP positive cells increased markedly in B and C groups.Conclusion:Human BMSCs possesse the potential of self renewing and proliferating,SHH can induce human BMSCs into the neural-like and/or glia-like cells under certain concantration.