南方医科大学学报

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (05): 513-516.

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成人骨髓间充质干细胞分化为神经元样细胞的逆转化现象

姚晓黎1, 张成1, 冯善伟1, 冯慧宇1, 刘祖国2, 邓宇斌3   

  1. 1. 中山大学第一医院神经科, 广东, 广州, 510080;
    2. 中山大学眼科中心, 广东, 广州, 510080;
    3. 中山大学病理生理教研室, 广东, 广州, 510080
  • 出版日期:2005-05-20 发布日期:2005-05-20
  • 基金资助:
    收稿日期:2004-10-6。
    基金项目:卫生部临床学科重点项目(2001321);广东省科技计划项目(2KM05501S);广东省科技计划项目重大专项(2003A3020106);国家自然科学基金(30270732)
    作者简介:姚晓黎(1971- ),女,博士,讲师,主要研究方向为干细胞移植治疗神经变性疾病和肌肉疾病,电话:020-33064439,020-87699289
    通讯作者:张成,电话:020-87335240,E-mail:czyrn@gzsums.edu.cn

Gene reversion of induced differentiation of adult human bone marrow-derived mesenchymal stem cells into neuron-like cells

YAO Xiao-li1, ZHANG Cheng1, FENG Shan-wei1, FENG Hui-yu1, LIU Zu-guo2, Deng Yu-bin3   

  1. 1. 中山大学第一医院神经科, 广东, 广州, 510080;
    2. 中山大学眼科中心, 广东, 广州, 510080;
    3. 中山大学病理生理教研室, 广东, 广州, 510080
  • Online:2005-05-20 Published:2005-05-20

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摘要： 目的 研究成人骨髓间充质干细胞(hMSC)分化为神经元样细胞后的逆转化现象.方法 从成人骨髓分离、培养和扩增hMSC.用参芪液诱导hMSC分化为神经元样细胞,并用免疫组化方法鉴定神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF)、胶质纤维酸性蛋白(GFAP)的表达.采用RT-PCR方法检测10个基因(内胚层基因ceruloplasmin;中胚层基因SM22:外胚层基因Amyloid precursor protein、syntaxin;生殖系基因protamine;神经元特异性基因Neuro D、NF、NSE、GFAP、Tau)在诱导分化为神经元样细胞的动态变化以及逆转化细胞的基因表达变化.结果 hMSC经参芪液诱导后,可见神经元样细胞.免疫组化显示诱导出的神经元样细胞表达NSE、NF阳性,GFAP阴性.去除参芪液,观察到神经元样细胞又恢复为hMSC的外形,呈现宽大扁平或长梭形.基因检测显示逆转化的细胞基因表达与未分化hMSC相似.结论 参芪液可以促进hMSC分化为神经元样细胞,诱导分化过程可以出现逆转化现象.

Abstract: Objective To observe gene reversion during differentiation of human adult bone marrow-derived mesenchymal stem cells (MSCs) into neuron-like cells induced by Shenqiye.Methods The MSCs were separated, cultured and expanded in the culture medium and induced to differentiate into neuron-like cells with Shenqiye. The expressions of neuron-specific enolase (NSE), neurofilament (NF), and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method, and the changes of 10 genes of the cells after differentiation were detected by reverse transcriptional PCR.Results The MSCs exhibited neuronal phenotype when treated with Shenqiye and the neuron-like cells were positive for expressions of NSE and NF but not for glial astrocyte marker GFAP. After withdrawal of Shenqiye from the medium, the neuron-like cells were reversed to MSC, flat or spindal in morphology. The gene expression profiles of the redifferentiated cells were similar to those of undifferentiated MSCs. Conclusion Shenqiye can induce MSC differentiation into neuron-like cells during and after which gene reversion may occur.

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