成人骨髓基质干细胞的体外培养及初步诱导分化研究  

A study on the culturing of human bone marrow stromal cells in vitro and its primary induction

作　　者：胡学昱[1] 罗卓荆[1] 田爽[2] 

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

出　　处：《中国脊柱脊髓杂志》2005年第10期594-597,i0003,共5页Chinese Journal of Spine and Spinal Cord

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

摘　　要：目的:探讨体外培养人骨髓基质干细胞(BMSC)的方法和初步诱导BMSC向神经细胞方向分化的方法。方法:采用正常成人献髓者骨髓,分离扩增BMSC,原代培养后将传1～4代细胞按1×104/ml种于24孔板,绘制生长曲线、贴壁率,观察细胞生长及不同浓度的碱性成纤维生长因子(bFGF)对BMSC的作用。以全反式维甲酸(RA)和bFGF为诱导剂,观察诱导前后BMSC的变化。结果:原代BMSC生长状态良好,传至第5代仍保持干细胞特性,bFGF可明显促进BMSC增殖,且呈剂量依赖关系。RA和bFGF诱导12h,BMSC逐渐向神经样细胞转化,胞体收缩成锥形、三角形或不规则形,细胞伸出细长突起,免疫细胞化学鉴定呈Nestin阴性,NSE阳性,GFAP阳性,且NSE阳性细胞数较GFAP阳性为少。结论:BMSC可在体外稳定扩增,且能保持干细胞特性,RA和bFGF可诱导其分化为神经细胞。Objective：To investigate the biological activity of human BMSC and its culturing method and explore the possibility of BMSC＇s differentiation into neurons in vitro.Method：BMSC were harvested from normal human marrow,then separated and proliferated.After primary cuhure,the subcuhured cells were implanted into hole plate by the number of 1×10^4/ml.The growth curve and maximum attached rate were described.The proliferating rate of BMSC were measured by basic fibroblast growth factor（bFGF） as inducer at different concentrations.BMSC were then cultivated in a medium containing all-transretinoic acid（RA） and bFGF.Microscopic observation,immunohistochemieal staining of Nestin,NSE and GFAP were performed.Result：The biological activity of primary cells sustained well,the subcuhured cells still kept the feature of BMSC evidently and showed the dose-effect relationship.NSE and GFAP positive cells were found after 14 days induction.Conclusion：BMSC can be cultivated automatically and proliferate stably ex vivo which indicates BMSC could be induced into neurons and glial cells in vitro.

关 键 词：骨髓基质干细胞 神经细胞 诱导分化 

分 类 号：Q813.1[生物学—生物工程] R329.2[医药卫生—人体解剖和组织胚胎学]