去分化肌肉干细胞神经增殖与分化能力的研究

Study of the neural proliferation and differentiation potential of dedifferentiated muscle stem cells

目的研究和鉴定去分化肌肉干细胞是否具有神经增殖和分化能力。方法使用不同的条件性培养基,对去分化肌肉干细胞依次进行神经增殖和神经分化诱导,并通过形态学、免疫细胞荧光化学、反转录聚合酶链式反应(RT-PCR)等手段加以鉴定。结果①去分化肌肉干细胞在神经增殖培养基诱导下,形成典型神经球(neurospheres)结构,5-乙炔基-2'脱氧尿嘧啶核苷(EdU)标记阳性,抗巢蛋白(Nestin)阳性;RT-PCR提示肌形成蛋白(myogenin)表达水平下调,而Nestin、干细胞抗原-1(Sca-1)表达上调。②经神经分化培养基诱导,神经球细胞可分化为形态学上典型的、抗中间神经丝蛋白(NFm)阳性神经元。结论去分化肌肉干细胞具有神经增殖和分化潜能。

Objective To investigate the ability of dedifferentiated muscle stem cells （De-MuSCs） to proliferate and differentiate along neural linenge by conditional cultivation. Methods De-MuSCs were cultured in neural proliferative media and then transferred into neural differentiation media for neural induction. Morphological observation, iranmnneytochemistry, 5-ethynyl-2-deoxyuridine （EdU） proliferation assay and reverse transcription- polymerase chain reaction （RT-PCR） analysis were performed for detection and evaluation. Results Neurosphere- like structures were observed when De-MuSCs cultured in neural proliferative media. Anti-Nestin staining combined with EdU detection showed the neural phenotype and proliferative status of the sphere cells that derived from De- MuSCs. RT-PCR explored the alteration of the gene expression of Nestin, stem cell antigen-1 （Sca-1） and myogenin during induction. After induction within neural differentiation media, some cells dissociated from the spheres were positive for Neurofilament-m （NFm） and displayed neuronal morphology. Conclusion De-MuSCs have the potential to conrnit to neural lineage and thus they are beneficial for neural regeneration.