脑源性神经生长因子基因转染对脊髓神经干细胞分化的影响

Differentiation of spinal cord derived neural stem cells modified by BDNF gene

目的 研究BDNF基因转染小鼠脊髓源性NSCs向神经元分化情况。方法 选取体外培养E14小鼠胚胎脊髓来源NSCs，构建整合有BDNF基因的逆转录病毒载体，感染体外培养的NSCs，诱导其向神经元分化。采用免疫细胞化学方法鉴定，确定NSCs的分化比例。结果 转染后诱导分化24h后可见部分细胞贴壁分化，48h后转染细胞大部分贴壁。BDNF转染NSCs分化为神经元比例较未转染NSCs明显增高，差异有统计学意义（P〈0．05）。结论 逆转录病毒载体介导BDNF基因转染NSCs可促进细胞分化，且分化多为神经元方向。

Objective To study the differentiation potential of mouse spinal cord derived neural stem cells （NSCs） into neurons after being transfected with BDNF gene in vitro. Methods Spinal cord derived NSCs from the El4 fetus mouse were isolated and cultured in vitro; the retrovirus containing pLXSN-BDNF gene was established and transfected into the above NSCs, and then, spinal cord derived NSCs were induced to be differentiated into neuron-like cells. Immunohistochemistry was employed to detect and calculate the ratio of differentiation of NSCs into neurons. Results The NSCs cultured in vitro partly adhered to the wall and differentiated within 24 h of transfection with BDNF gene, and most of the NSCs adhered to the wall differentiated within 48 h oftransfection. The level of neurons from spinal cord derived NSCs modified by BDNF gene was markedly increased as compared with that that from normal spinal cord derived NSCs （P〈0.05）. Conclusion NSCs transfected by retroviral pLXSN-BDNF can promote the cell differentiation. BDNF gene can increase greatly the percentage of neurons in the course of inducing the differentiation of mouse NSCs.