中国组织工程研究2018，Vol.22Issue(17)：2692-2698,7.DOI:10.3969/j.issn.2095-4344.0482
鼠神经生长因子联合脑源性神经营养因子诱导人脐血间充质干细胞向神经元样细胞分化
Human umbilical cord blood-derived mesenchymal stem cells differentiate into neuron-like cells induced by combination of mouse nerve growth factor and brain-derived neurotrophic factor
摘要
Abstract
BACKGROUND: We have found that mouse nerve growth factor has the ability to induce differentiation of umbilical cord blood mesenchymal stem cells (UCB-MSCs) into neurons in vitro. In order to further explore the method of improving the induction efficiency of nerve cells, we attempt to combine a variety of cell growth factors for cell induction. OBJECTIVE: To explore the effect of mouse nerve growth factor combined with brain-derived neurotrophic factor on the differentiation of UCB-MSCs into neuron-like cells in vitro . METHODS: After the donated primary UCB-MSCs were resuscitated and cultured, the passage 5 UCB-MSCs were divided into five groups. The first four groups served as pre-induced groups, and fibroblast growth factor and epidermal growth factor were added to pre-induce cells for 24 hours, and mouse nerve growth factor and brain-derived neurotrophic factor, alone or in combination, were used thereafter to induce UCB-MSCs, while in control group, only the same amount of cell medium was added. The last group was non-pre-induced group, in which the cells were cultured in the cell culture medium for 24 hours, and then mouse nerve growth factor and brain-derived neurotrophic factor were both added to induce UCB-MSCs. The morphological changes of cells were observed under inverted microscope. The expression of neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) was detected by immunocytochemistry technique. Real-time qPCR was used to detect the relative expression of NSE and GFAP at mRNA level. RESULTS AND CONCLUSION: (1) The cell morphology of UCB-MSCs was in long shuttle shape and spindle shape with unequal size. After induction, the cell bodies gradually retracted and became rounded, and the projections extended to one-side or multi-sides, presenting with the neuron-like changes. (2) Immunocytochemistry and real-time qPCR results showed that NSE and GFAP were positive in each experimental group, and the positive rate and mRNA expression of NSE and GFAP in the combined induction group were higher than those in the other groups. (3) Either mouse nerve growth factor or brain-derived neurotrophic factor could induce UCB-MSCs to differentiate into neuron-like cells. Moreover, there was a cumulative effect between the two cytokines, and their combined use could effectively improve the efficiency of induction.关键词
干细胞/脐血/间充质干细胞/鼠神经生长因子/脑源性神经营养因子/神经元样细胞/诱导分化分类
医药卫生引用本文复制引用
陈俊,杨自金,李红梅..鼠神经生长因子联合脑源性神经营养因子诱导人脐血间充质干细胞向神经元样细胞分化[J].中国组织工程研究,2018,22(17):2692-2698,7.基金项目
江苏省连云港市科技局重点科室建设(SH1117) the Key Division Construction of Lianyungang Municipal Science and Technology Department, No. SH1117 （SH1117）
