脑源性神经营养因子基因转染骨髓间充质干细胞体外诱导分化为神经元样细胞初步观察.pdfVIP

252 Journal of Audíology and Speech Pathology 2009. Vo117. No. 3 ·实验研究· 脑源性神经营养因子基因转染骨髓间充质干细胞 体外诱导分化为神经元样细胞初步观察祷 刘谦虚l 谢鼎华1 陈观贵1 【摘要 阔的 探讨骨髓间充质子细胞( bone …marrow mesenchymal stem cells , BMSCs)转娩人脑源性神经 营养因子(braín…d盯íved neurotrophic factor. BDNF) 基因后在体外分化为神经元样细胞的功能。穷法 克除人 BDNF 基因并构建其真核表达载体;分离培养 5 只豚鼠 BMSCs.观调~其形态并用流式细胞仪鉴定$将人 BDNF 基 因电穿孔法转染 BM仪坏.G418 筛选后用维阳酸诱导分化，免疫细胞化学法对分化细胞进行鉴定。结果 分离培养 的细胞具有典型的 BMSCs 形态和标志，转染诱导盾的细胞表达神经元特异性烯蹲化酶、巢萤臼(Nestín) 和胶质纤 维酸性蛋白，并能分泌 BDNF 。结论 BDNF 葱因转染的 BMSCs 在体外能分化为神经元样细胞，电穿孔法能提高 转染率.RA 有促诱导作用。 E关键词】 脑源性神经营养因子慕因; 骨髓间充质干细胞s 诱导分化s 神经元样细胞 E 中倒分类号】 R339.16 【文献标识码】 A (父黛编号] 1006-7299(2009)03一0252 …04 A Preliminary Study on the Differentiation of Bone … marrow Mesenchymal Stem Oells into Neuron-like Cells in Vitro after Transfected by Brain- derived Neurotrophic 俨actor Gene Liu Qianxu.Xie Dinghua , Chen Guangui (Hearing Research Institute.Department of Otolaryngology and Head Neck Surgery of the Second Xiangya Hospital , Central South University. Changsha , 410011 ,Ohina) (Abstract Objective To investigate the function of bone … marrow me阴nchymal stem cells( BMSCs) differ- entiating ínto neuron 一 líke cells ín vitro after transfected by human braín - deríved neurotrophíc factor (BDNF) gene. Methods Human BDNF genes were cloned and recombinant pcDNA3. l(…)… BDNF plasmíds were construc- ted. BMSCs from five guínea pígs were ísolated and cultured while their morphologies were observed by microscope. The surface antigen was detected by flowcytometry. BDNF genes were transfected into BMSCs with electroporation. and the transfected BMSCs were induced by ratinoi