脑源性神经营养因子体外诱导骨髓间充质干细胞分化为神经元样细胞

南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (08): 854-858.

脑源性神经营养因子体外诱导骨髓间充质干细胞分化为神经元样细胞

黄文, 张成, 陈松林, 张为西, 姚晓黎, 曾缨, 黄慧, 冯善伟, 柳太云

中山大学医学院附属第一医院神经内科, 广东, 广州, 510080

出版日期:2004-08-20 发布日期:2004-08-20
基金资助:
收稿日期:2004-3-19。
基金项目:国家自然科学基金资助项目(30170337,30370510);广东省自然科学基金资助项目(31693);教育部骨干教师基金资助项目(20002349);卫生部临床学科重点项目资助(2001321);教育部博士点资金资助(20030558058)
作者简介:黄文(1968-),女,广西灵山人,2004年博士毕业于中山大学附属第一医院,副教授,现在广西医科大学附属第一医院神经科,研究方向为神经肌肉遗传病,电话:0771-5356735,E-mail:hwenl229@yahoo.com.cn
通讯作者:张成,电话:020-87332387,E-mail:czym@gzsums.edu.cn

Brain-derived neurotrophic factor induces rat bone marrow stromal cells to differentiate into neuron-like cells in vitro

HUANG Wen, ZHANG Cheng, CHEN Song-lin, ZHANG Wei-xi, YAO Xiao-li, ZENG Ying, HUANG Hui, FENG Shan-wei, LIU Tai-yun

中山大学医学院附属第一医院神经内科, 广东, 广州, 510080

Online:2004-08-20 Published:2004-08-20

摘要/Abstract

摘要： 目的探讨脑源性神经营养因子(BDNF)体外诱导骨髓间充质干细胞(MSCs)分化成神经元样细胞的作用及其对诱导后的神经元样细胞保护作用,为治疗神经系统疾病提供更优良的细胞。方法体外培养MSCs至第5代后,分别用BDNF、β-巯基乙醇(β-ME)诱导MSCs,诱导后的第1、3和6小时计算两组神经元样细胞数,对各时间点两组神经元样细胞阳性率进行了比较;诱导后行神经细胞标记抗体的免疫细胞化学、RT-PCR及蛋白质印迹鉴定。结果两组诱导剂诱导后的细胞均呈神经元样细胞的形态;BDNF组诱导分化后细胞的存活时间远较β-ME组长;BDNF组在诱导后6h近于诱导高峰,而β-ME在诱导2h即达高峰,但随后神经元样细胞渐死亡。免疫细胞化学结果显示在两组诱导后细胞的巢蛋白(Nestin)、神经细胞特异性烯醇化酶(NSE)、微管相关蛋白-2(MAP-2)、神经丝蛋白(NF)表达均呈阳性,而胶质纤维酸性蛋白(GFAP)表达呈阴性;RT-PCR显示NSE、NFL,MAP-2的mRNA表达阳性,GFAP有较弱的表达;蛋白质印迹可见两组诱导剂诱导的细胞均表达神经元的特异性标记抗体NSE。结论 BDNF能单独诱导MSCs分化形成神经元样细胞,而且分化后细胞的存活时间长,有望用于治疗脑缺血、神经变性等疾病。

Abstract: Objective To investigate brain-derived neurotrophic factor (BDNF)-induced differentiation of rat bone marrow stromal cells (MSCs) into neuron-like cells in vitro and observe its neuroprotective effect of BNDF on the differentiated cells, which might provide the better seed cells for treatment of nervous system diseases. Methods The fifth-passage MSCs were induced by BDNF and 2-mercapto ethanol(β-ME)respectively, 1, 3 and 6 h after which the induced neuron-like cells were counted and compared. At 3 h, the neuron-like cells were identified by the immunocytochemical staining, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting. Results The two induced cells both displayed neuronal morphologies with long and multipolar cell projections, but BDNF-induced cells survived for a longer time than γ-ME-induced ones. The results of immunocytochemical staining showed that the two neuron-like cells expressed nestin, neuron-specific enolase (NSE), neurofilament (NF), microtubule-associated protein (MAP-2), but not glial fibrillary acidic protein (GFAP). RT-PCR detected mRNA-positive NSE, NF and MAP-2 in the induced cells, with also mild positive GFAP mRNA. Western blotting identified also NSE expression in these neuron-like cells. Conclusion BDNF alone may induce rat MSCs to differentiate into neuron-like cells in vitro, which have longer lifetime to better serve the purpose of transplantation and gene therapy for nervous system diseases.

中图分类号:

R338.8

黄文, 张成, 陈松林, 张为西, 姚晓黎, 曾缨, 黄慧, 冯善伟, 柳太云. 脑源性神经营养因子体外诱导骨髓间充质干细胞分化为神经元样细胞[J]. 南方医科大学学报, 2004, 24(08): 854-858.

HUANG Wen, ZHANG Cheng, CHEN Song-lin, ZHANG Wei-xi, YAO Xiao-li, ZENG Ying, HUANG Hui, FENG Shan-wei, LIU Tai-yun. Brain-derived neurotrophic factor induces rat bone marrow stromal cells to differentiate into neuron-like cells in vitro[J]. Journal of Southern Medical University, 2004, 24(08): 854-858.

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