神经生长因子转染骨髓间充质干细胞诱导分化成神经样细胞

目的探讨神经生长因子(NGF)转染大鼠骨髓间充质干细胞(MSCs)后的表达及其对MSCs分化成神经样细胞的影响。方法在体外低密度扩增大鼠骨髓MSCs。应用基因重组技术,构建pEGFP-NGF真核表达质粒,并将其转染至MSCs中。免疫荧光标记法检测中间神经丝蛋白(NF-M)和胶质纤维酸性蛋白(GFAP)的表达。在荧光显微镜下观察MSCs阳性荧光的表达率、细胞的形态变化和类型。结果MSCs阳性荧光的表达率约为30%。转染后,MSCs呈现多突起,且几个突起之间互相连接成网状,似神经元样的形态。免疫荧光标记法鉴定其中一部分细胞表达NF-M,另一部分细胞表达GFAP。结论转染的MSCs可表达NGF并在含有NGF的微环境中分化成神经样细胞。

Mesenchymal stem cells of bone marrow differentiate into neuron-like cells after transfected by nerve growth factor gene in vitro

Objective To study the gene expression of nerve growth factor(NGF) in rat bone marrow mesenchymal stem cells(MSCs)and the effect of NGF-GFP fusion protein on MSCs differentiation into neuron-like cells.Methods Rat bone marrow mesenchymal stem cells were amplificated in low density in vitro。The NGF gene was inserted into pEGFP-N2C-terminal protein fusion vector to construct pEGFP-NGF,which was transfected into MSCs using Lipofectamine 2000.The infected efficiency,morphological change of MSCs and cell types were examined by fluorescent microscope.Results Infected efficiency of NGF-GFP fusion protein was 30% in MSCs.The transfected MSCs displayed neuron-like morphology and formed net.Some transfected MSCs expressed neurofilament protein-M(NF-M),while the others was immuostained by glial fibrillary acidic protein(GFAP).Conclusion MSCs might be used as carrier cells of gene therapy because they express NGF and differentiate into neuron-like cells in the environment with NGF.