Sox2 induce mouse embryonic fibroblasts direct reprogramming into neural stem cells |
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| Authors: | LIU Chang RONG Li-min LIU Bin |
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| Institution: | Department of Spine Surgery, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China |
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| Abstract: | AIM: To study the direct reprogramming method of mouse embryonic fibroblasts (MEFs) converted into induced neural stem cells (iNSCs). METHODS: Sox2-infected MEFs were cultured in NSCs culture medium for 10 d. Subsequently, repeated suspension and adherent culture were performed for 3 times for the purification of iNSCs. The iNSCs were cultured in suspension medium. Real-time PCR was used to detect the expression of neural stem cell marker genes and pluripotent marker gene. In vivo, iNSCs were microinjected into the mouse cerebral cortex. Immunofluorescence was performed to detect the expression of neural stem cell, neuron, oligodendrocyte and astrocyte markers in vitro and vivo. RESULTS: A variety of neural stem cell marker gene expression was significantly increased in iNSCs detected by real-time PCR. Immunofluorescence confirmed that iNSCs expressed nestin and differentiated into neurons, oligodendrocytes and astrocytes in vitro and vivo. CONCLUSION: Sox2 is sufficient to trigger the direct reprogramming from MEFs to iNSCs. iNSCs have the ability of self-renew and 3 differentiation potentials in vivo and vitro. iNSCs are the suitable seed cells of SCI. |
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| Keywords: | Sox2 Mouse embryonic fibroblasts Direct reprogramming Neural stem cells |
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