丹参酮ⅡA定向诱导骨髓间质干细胞分化为神经元样细胞的研究  

Human mesenchymal stem cells differentiate into neuron-like cells with Tanshinone II A

作　　者：夏文杰[1] 项鹏[1] 张丽蓉[2] 陈振光[3] 余伟华[1] 张秀明[1] 李艳[1] 李树浓[1] 

机构地区：[1]中山大学中山医学院病理生理学教研室,广东广州510080 [2]广东药学院病理生理学教研室,广东广州510224 [3]中山大学附属第一医院心胸外科,广东广州510080

出　　处：《中国病理生理杂志》2003年第7期865-869,T001,共6页Chinese Journal of Pathophysiology

基　　金：国家重点研究发展规划项目 ( 973) (No .2 0 0 1CB5 0 990 4 );广东省十五专项 (A30 2 0 10 1);国家自然科学基金资助项目 ( 30 10 0 188)

摘　　要：目的 :丹参酮ⅡA体外定向诱导成人骨髓间质干细胞 (MSC)分化为神经元样细胞。方法 :采用Fi coll-Paque液离心和贴壁法分离成人MSC ,体外扩增 ,流式细胞仪检测MSC表面抗原表达 ,FGF - 2对MSC增殖和分化的影响。采用含丹参酮ⅡA的无血清达乐伯克改良必需基本培养基 (DMEM)诱导MSC分化为神经元样细胞 ,免疫组化鉴定神经元特异性烯醇化酶 (NSE)、神经丝蛋白 (NF -M)、胶质纤维酸性蛋白 (GFAP)、巢蛋白 (nestin)的表达。结果 :成人骨髓间质干细胞在体外扩增原代可获得 0 5× 10 6,15代可获得 ( 2 - 3)× 10 12 个细胞 ,流式细胞仪检测结果显示CD2 9、CD4 4、CD90、CD10 5、CD16 6表达阳性 ,CD11a、CD14、CD34、CD38、CD4 5、CD80、CD86为阴性。FGF - 2促进hMSC生长 ,并且对MSC的分化能力基本没有影响。丹参酮ⅡA诱导后 ,MSC胞体收缩 ,突起伸出 ;免疫组化显示诱导出的神经元样细胞NSE、NF -M、nestin表达阳性 ,GFAP阴性。结论AIM:To investigate the differentiation from human mesenchymal stem cells(hMSC) into neuron-like cells with Tanshinone II A. METHODS: hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. To detect the surface antigens, the labeled cells were analysed on a FACScan flow cytometer to determine the effect of the capacity of proliferation and differentiation of the mesenchymal stem cells with FGF-2. hMSC were induced to differentiate into neurons with DMEM Tanshinone II A. Neuron-specific enolase(NSE), neurofilament(NF), Nestin, glial fibrillary acaidic protein(GFAP) were detected by immunohistochemistry. RESULTS: hMSC were expanded as undifferentiated cells in culture for more than 15 passages. The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. These expanded attached MSC were uniformly positive for CD29,CD44,CD90,CD105,CD166 and didn't express CD11a ,CD14,CD34,CD38,CD45,CD80,CD86. FGF-2 have special effect on low denisity MSCs. Simple methods with Tanshinone II A induced hMSC to exhibit a neuronal phenotype, expressing NSE, NF-M, Nestin at 5 hours. But the neuron-like cells didn't express the glial astrocyte marker GFAP. CONCLUSION: hMSC can be induced to differentiate into neurons with Tanshinone II A.

关 键 词：骨髓 间质干细胞 神经元 丹参酮ⅡA 成纤维细胞生长因子 细胞分化 

分 类 号：R338.8[医药卫生—人体生理学]