去分化肌肉干细胞神经分化潜能的探讨

Investigation of dedifferentiated muscle stem cells committing to neural linage

目的探讨去分化肌肉干细胞经条件培养诱导,具有神经干细胞性质并分化为终末神经细胞的潜能。方法依次用神经干细胞增殖及神经细胞分化条件培养基,对去分化肌肉干细胞进行体外诱导,促使其向神经干细胞转变以及向终末神经细胞分化,并通过形态学、免疫细胞化学、RT-PCR等实验手段研究其性质并加以鉴定。结果 (1)去分化肌肉干细胞在神经干细胞增殖条件培养基诱导下,可形成神经球,EdU标记阳性,抗Nestin、Neurofilament-m(NFm)、GFAP、CNPase阳性;Myogenin表达水平下调,而Nestin、Sca-1表达上调;(2)经神经细胞分化条件培养基诱导,神经球细胞可分化为形态学上典型的、抗NFm阳性神经元,以及抗GFAP、CNPase阳性神经胶质细胞。结论去分化肌肉干细胞具有神经系的多分化潜能。

Objective To investigate the ability of dedifferentiated muscle stem cells（De-MuSCs） to commit to neural lineage by conditional cultivation.Methods De-MuSCs were cultured in neural stem cell（NSC） proliferative media for neurosphere formation and then transferred into neural differentiation（ND） media for neuronal induction.Morphological observation,immunocytochemistry,EdU proliferation assay and RT-PCR analysis were performed for the detection and evaluation.Results（1）Neurosphere-like structures were observed when De-MuSCs were cultured in NSC proliferative media.Anti-nestin,neurofilament-m（NFm）,GFAP and CNPase staining combined with EdU detection showed the neural phenotype and proliferative status of the cells in the spheres that derived from De-MuSCs.RT-PCR explored the alteration of the gene expression of Nestin,Sca-1 and myogenin during the induction.（2）After induction within ND media,some cells dissociated from the spheres were solely positive for NFm and possessed long and thin projections that resembled neuronal processes.GFAP or CNPase positive cells were observed as well.Conclusion De-MuSCs have the potential to commit to neural lineage and may be able to aid recovery from neurological diseases by providing a new competitive cellular source for neural regeneration.