CAJ | 학술논문

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。
배경：Neurogenin2(Ngn2)기인조공성형효질세포분화위신경원이피실험증실，저제시허왕세포야가능통과기인조공분화위신경원。 　　목적：연구Neurogenin2기인조공대서허왕세포향신경원분화적가행성。 　　방법：체외배양、순화급감정대서허왕세포，연후용함록색형광단백기인적만병독재체계통장Neurogenin2기인전염도입허왕세포중，최후용함감성성섬유세포생장인자、표피생장인자화뇌원성세포생장인자적무혈청DMEM유도배양허왕세포2주。현미경관찰세포형태，면역세포화학검측수린지감성단백급신경원특이성희순화매。 　　결과여결론：허왕세포전염도입 Neurogenin2기인병유도분화후，면역형광검측발현12.56%적세포표체신경원표지성단백신경원특이성희순화매，이대조조균미발현표체신경원표지성단백신경원특이성희순화매。Neurogenin2기인식입가사대서허왕세포표체신경원표지성단백신경원특이성희순화매，제시해기인가조공허왕세포전분화위신경원。
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.