The study on preparation of reprogrammed neural stem cells derived from skin and neural differentiation in vitro
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    Abstract:

    Objective:To evaluate the feasibility of preparation of reprogrammed neural stem cells derived from skin and neural differentiation in vitro. Methods:Skin-drived precursors (SKPs) were isolated and purified from rat skin tissue,identified and seeded at passage 3 in 12-well culture plates. SKPs were divided into three groups:group A was transfected with neurogenin2 via GFP-plentivirus;group B was transfected with GFP-plentivirus;group C was cultured without transfection. All groups were induced by common inducer for a week after 7 days of transfection.The morphology of induced SKPs was observed by microscope,and the markers of neural cells(MAP-2 and NeuN) were detected by immunocytochemistry. Results:The green fluorescence of SKPs in group A reached the peak on the 7th day after transfection and existed until the 8th passage.The shapes of SKPs were fusiform or ellipse, and most SKPs express several apophyses after inducement. Immunofluorescence demonstrated that 90.98% of the SKPs expressed MAP-2 and 62.23% of the SKPs expressed NeuN. SKPs in group B expressed green fluorescence and had no significant typical morphological changes after inducement. Immunofluorescence demonstrated that 34.35% of the SKPs epressed Map-2 and no SKPs expressed NeuN;SKPs in group C did not express green fluorescence and had no significant typical morphological changes after inducement. Immunofluorescence demonstrated that 29.54% of the SKPs expressed MAP-2 and no SKPs expressed NeuN. Conclusion:The reprogrammed neural stem cells can be prepared through transfecting neurogenin2 into SKPs via plentivirus in vitro, and their capability of differentiating into neural cells elevated visibly after neurogenin 2 transfection.

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代学良,程 峰,郝怀勇,钱腾达,李立新.皮肤源性再程序化神经干细胞的制备以及定向分化为神经细胞的体外研究[J].南京医科大学学报(自然科学版英文版),2011,(3):319-323.

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  • Received:September 10,2010
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