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

作　　者：唐寅达[1] Wen Xu Xiaodong Mu Yong Li 李世亭[1] 

机构地区：[1]上海交通大学医学院附属新华医院神经外科,上海200092 [2]匹兹堡大学医学院附属儿童医院干细胞研究中心,美国匹兹堡15213 [3]德克萨斯大学休斯顿医学中心布朗分子医学研究所干细胞与再生医学研究中心,美国休斯顿77030

出　　处：《中华神经外科疾病研究杂志》2013年第3期222-226,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：上海市科协技术委员会科研计划国际合作基金资助项目(10410711400)

摘　　要：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.

关 键 词：去分化 去分化肌肉干细胞 神经干细胞 神经分化 细胞培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]