成人骨髓基质细胞体外诱导成神经细胞实验研究  

Induction of human bone marrow stromal cells into neurons and glial cells in vitro

作　　者：胡学昱[1] 罗卓荆[1] 田爽[2] 宫伟[1] 褚尤彪[1] 刘艳武[1] 胡慧敏[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,陕西西安710032 [2]第四军医大学西京医院西京医院妇产科,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2006年第2期139-142,共4页Chinese Journal of Neurosurgical Disease Research

基　　金：国家863高技术研究发展计划基金资助项目(2002AA216101)

摘　　要：目的探讨骨髓基质细胞的体外培养、扩增及诱导其向神经元样细胞定向分化的方法。方法采用改进的全骨髓培养法,培养来源于正常成人献髓者的骨髓基质细胞。传至第5代,用全反式维甲酸(RA)等细胞因子诱导分化,于诱导后第7d和第14d行神经元细胞特异性抗原神经元特异性烯醇化酶(NSE)、微管相关蛋白-2(MAP-2)、神经丝蛋白(β-tublin)和胶质纤维酸性蛋白(GFAP)鉴定,计数诱导前后的细胞。结果成人骨髓基质细胞经RA+碱性成纤维细胞生长因子(bFGF)+脑源性神经营养因子(BDNF)诱导后可见神经元样细胞出现,经NSE、MAP-2及β-tublin免疫组化和荧光鉴定阳性,NSE表达率达到48.5%±0.2%。结论骨髓基质细胞在体外扩增迅速,纯化需时短,并可在全反式维甲酸等细胞因子诱导下向神经元样细胞分化。Objective To study the methods of culture, amplification of human bone marrow stromal cells （BMSCs）, and induction of BMSCs differentiating into the neuron-like cells in vitro. Methods The BMSCs were isolated primarily from bone marrow of adult, and purified by passage culture. The 5 th passage of BMSCs was induced by all-trans-retinoic acid （RA）. Immunocytochemical detection of neuron specific enolase （NSE）, protein 2 （MAP-2）, β-tublin, and glial fibrillary acidic protein（GFAP） were also performed on cells after 7 d and 14 d induction. And cell numbers pre- and post-induction were counted. Results Neuron-like cells appeared airing BMSCs after BMSCs induced by RA ＋ bFGF ＋ BDNF. Neuron-like cells were showed NSE, MAP-2 and β-tublin positive in immunohistochemical and immunofluorescence staining.The positive percentage of NSE protein expression was48.5% ±0.2%. Conclusion BMSCs can be amplified and purified in short time and be induced into neuronal-like cell by RA.

关 键 词：骨髓基质细胞 神经细胞 全反式维甲酸 分化 

分 类 号：R329.29[医药卫生—人体解剖和组织胚胎学]