【摘要】 目的:探讨胶质细胞源性神经营养因子(glial cell line-derived neurotrophic factor,GDNF)基因修饰的骨髓间充质干细胞在体外诱导分化为神经元样细胞的潜能。方法:密度梯度离心和贴壁培养相结合分离纯化大鼠骨髓间充质干细胞(BMSCs)并鉴定,分别用GDNF基因重组腺病毒(pAdEasy-1-pAdTrack-CMV-GDNF)和空质粒腺病毒(pAdEasy-1-pAdTrack-CMV)感染BMSCs建立BMSCs/GDNF和BMSCs/GFP两种细胞。两种细胞用DMSO/β-ME/ATRA进行诱导分化,观察细胞形态变化,采用Hoechst染色和免疫荧光细胞化学染色检测细胞凋亡和分化情况。结果:与BMSCs/GFP组相比,BMSCs/GDNF组的BMSCs经诱导后细胞凋亡率降低,NF-200阳性细胞率上升,但GFAP阳性细胞率无明显变化。结论:GDNF基因修饰有利于BMSCs抗细胞凋亡和分化为神经元样细胞,这为进一步研究GDNF基因修饰的BMSCs应用于中枢神经系统疾病的治疗奠定了基础。更多还原

【Abstract】 Objective: To explore the potency of GDNF gene modified bone marrow mesenchymal stem cells(BMSCs) to differentiate into neruon-like cells in vitro.Methods:BMSCs were separated and purified by density gradient centrifugation and cell cultivation;and BMSCs/GDNF and BMSCs/GFP cell lines were established by transfecting with pAdEasy-1-pAdTrack-CMV-GDNF and empty vector pAdEasy-1-pAdTrack-CMV.After inducing by DMSO/β-ME/ ATRA in vitro,the cell morphology were observed by microscope,apoptosis was analyzed by Hoechst staining,and cell differentiation were detected by immunofluorescence staining.Results:BMSCs apoptotic rate was repressed,and the rate of NF-200 positive cells was increased in BMSCs/GDNF group compared with BMSCs/GFP group,but the rate of GFAP positive cells had no significant change.Conclusion:GDNF gene-modification helps BMSCs to resist apoptosis and differentiate into neuron-like cells.更多还原