摘要
背景:Neurogenin2(Ngn2)基因调控星形胶质细胞分化为神经元已被实验证实,这提示许旺细胞也可能通过基因调控分化为神经元。目的:研究Neurogenin2基因调控大鼠许旺细胞向神经元分化的可行性。方法:体外培养、纯化及鉴定大鼠许旺细胞,然后用含绿色荧光蛋白基因的慢病毒载体系统将Neurogenin2基因转染导入许旺细胞中,最后用含碱性成纤维细胞生长因子、表皮生长因子和脑源性细胞生长因子的无血清DMEM诱导培养许旺细胞2周。显微镜观察细胞形态,免疫细胞化学检测髓磷脂碱性蛋白及神经元特异性烯醇化酶。结果与结论:许旺细胞转染导入Neurogenin2基因并诱导分化后,免疫荧光检测发现12.56%的细胞表达神经元标志性蛋白神经元特异性烯醇化酶,而对照组均未发现表达神经元标志性蛋白神经元特异性烯醇化酶。Neurogenin2基因植入可使大鼠许旺细胞表达神经元标志性蛋白神经元特异性烯醇化酶,提示该基因可调控许旺细胞转分化为神经元。
BACKGROUND：It is confirmed that astrocytes can differentiate into neurons by Neurogenin2 gene regulation, suggesting that Schwann cells may also differentiate into neurons by gene regulation.
OBJECTIVE：To evaluate the feasibility of Schwann cells differentiating into neurons by Neurogenin2 gene regulation.
METHODS：Rats Schwann cells were isolated, purified and identified. Then the Schwann cells were transfected with Neurogenin2 via green fluorescent protein gene-plentivirus. To induce neuronal differentiation, the Schwann cells were cultured in serum-free Dulbecco’s modified Eagle’s medium containing epidermal growth factor, basic fibroblast growth factor and brain-derived neurotrophic factor for 2 weeks. The morphology of induced cells was observed by microscope, and myelin basic protein and neuron-specific enolase were detected by immunocytochemistry.
RESULTS AND CONCLUSION：After transfection with Neurogenin2 via green fluorescent protein gene-plentivirus and induced differentiation, immunofluorescence assay demonstrated that 12.56%of the induced cellexpressed neuron-specific enolase, but the control group did not express neuron-specific enolase. Neurogenin2 gene-transfected Schwann cells can express neuron-specific enolase, suggesting Neurogenin2 gene may regulate transdifferentiation of Schwann cells into neurons.
出处
《中国组织工程研究》
CAS
CSCD
2013年第49期8590-8595,共6页
Chinese Journal of Tissue Engineering Research
基金
宁波市自然科学基金资助项目(2010A610070)~~
