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首页> 美国卫生研究院文献>Experimental and Therapeutic Medicine >Brain-derived neurotrophic factor gene-modified bone marrow mesenchymal stem cells
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Brain-derived neurotrophic factor gene-modified bone marrow mesenchymal stem cells

机译：脑源性神经营养因子基因修饰的骨髓间充质干细胞

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摘要

The present study aimed to investigate the effects of human brain-derived neurotrophic factor (hBDNF) on the differentiation of bone marrow mesenchymal stem cells (MSCs) into neuron-like cells. Lentiviral vectors carrying the hBDNF gene were used to modify the bone marrow stromal cells (BMSCs) of Sprague-Dawley (SD) rats. The rat BMSCs were isolated, cultured and identified. A lentivirus bearing hBDNF and enhanced green fluorescent protein (eGFP) genes was subcultured and used to infect the SD rat BMSCs. The expression of eGFP was observed under a fluorescence microscope to determine the infection rate and growth of the transfected cells. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) was used to detect the proliferation rate of cells following transfection. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used to detect the expression levels of hBDNF. Differentiation of neuron-like cells was induced in vitro and the differentiation rate of the induced neural-like cells was compared with that in control groups and analyzed statistically. In the cultured cells, flow cytometry demonstrated positive expression of cluster of differentiation (CD)90 and CD44, and negative expression of CD34 and CD45. The proliferation rate of the rat BMSCs increased following gene transfection. The expression of hBDNF-eGFP was detected in the BMSCs of the experimental group. The differentiation rate of hBDNF-modified cells into neuron-like cells in the experimental group was higher compared with that in empty plasmid and untransfected negative control groups. The difference was statistically significant (P<0.05). Thus, BDNF gene transfection is able to promote the differentiation of BMSCs into neuron-like cells. BDNF may play an important role in the differentiation of MSCs into neuron-like cells.
机译：本研究旨在研究人脑源性神经营养因子（hBDNF）对骨髓间充质干细胞（MSCs）分化为神经元样细胞的影响。携带hBDNF基因的慢病毒载体用于修饰Sprague-Dawley（SD）大鼠的骨髓基质细胞（BMSC）。分离，培养和鉴定大鼠BMSC。携带hBDNF和增强型绿色荧光蛋白（eGFP）基因的慢病毒被继代培养，并用于感染SD大鼠BMSC。在荧光显微镜下观察eGFP的表达，以确定感染率和转染细胞的生长。甲基噻唑基二苯基溴化四唑（MTT）用于检测转染后细胞的增殖速率。逆转录定量聚合酶链反应（RT-qPCR）和蛋白质印迹分析被用来检测hBDNF的表达水平。在体外诱导神经元样细胞的分化，并将诱导的神经样细胞的分化率与对照组进行比较并进行统计学分析。在培养的细胞中，流式细胞仪显示分化簇（CD）90和CD44阳性表达，而CD34和CD45阴性表达。基因转染后，大鼠骨髓间充质干细胞的增殖速率增加。在实验组的骨髓间充质干细胞中检测到hBDNF-eGFP的表达。实验组hBDNF修饰细胞向神经元样细胞的分化率高于空质粒和未转染的阴性对照组。差异具有统计学意义（P <0.05）。因此，BDNF基因转染能够促进BMSCs向神经元样细胞的分化。 BDNF可能在MSC分化为神经元样细胞中起重要作用。

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