摘要
目的研究神经营养素-3(neurotrophin-3,NT-3)对骨髓间充质干细胞(mesenchymal stem cells,MSCs)增殖和向神经元分化的影响,探讨转染NT-3基因的MSCs体内移植治疗脊髓损伤的可行性。方法体外培养稳定表达NT-3蛋白的MSCs,通过MTT法检测NT-3对MSCs增殖的影响;应用免疫细胞化学和免疫印迹方法检测转染NT-3的MSCs表达神经元标志物NeuN蛋白的情况。结果转染NT-3基因的MSCs生长曲线上移,细胞增殖率显著升高,与对照组相比,两者有显著差异(P<0.05),表明转染NT-3后细胞生长加快。免疫荧光染色结果显示转染NT-3的MSCs的NeuN阳性细胞数明显增多,与对照组比较有显著差异(P<0.05);Western印迹检测结果显示转染NT-3的MSCs内NeuN蛋白表达明显增多,与对照组比较有显著差异(P<0.05),表明NT-3促进MSCs向神经元方向分化。结论 NT-3基因转染可促进MSCs的增殖,并诱导其向神经元方向分化。
Objective To study the differentiation potential of bone mesenchymal stem cells（MSCs） into neurons after being transfected with neurotrophin-3（NT-3） gene in vitro.Methods The MSCs stablely expressed NT-3 protein was cultured in vitro,and the effect of proliferation of NT-3 on MSCs was detected by MTT assay.The expression of NeuN protein in MSCs transfected with NT-3 gene was detected by immunocytochemistry.Results The growth curve of gene moved up,the ratio of cell proliferation was significantly rised,there was significant difference between two groups（P0.05）.It indicated that the cells grow was accelerated.More NeuN-positive cells in MSCs were transfected with NT-3 compared with those of control group（P0.05）.Western blot assay indicated that the expression of NeuN protein in NT-3 transfected MSCs was significantly different compared with that of control.It demonstrated that NT-3 promoted MSCs to differentiate into neuron-like cells.Conclusions The transfection with NT-3 in MSCs could promote proliferation of MSCs and differentiate them into neurons-like cells.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2012年第8期1624-1626,共3页
Chinese Journal of Gerontology
基金
国家自然科学基金(81171013)
徐州市科技发展基金(XF10C079)
徐州医学院院长人才专项基金(08KJZ04)
