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作 者:余勤[1] 罗依[2] 董勤[3] 赵伟春[1] 刘文洪[1] 孙国金[1]
机构地区:[1]浙江中医学院生物工程学系,杭州310053 [2]浙江大学医学院附属第一医院血液科 [3]浙江省中医院儿科
出 处:《中华急诊医学杂志》2004年第12期826-829,i004,共5页Chinese Journal of Emergency Medicine
基 金:浙江省自然科学基金资助项目 (M3 0 3 70 7);浙江省科技厅资助项目 (2 0 0 4C3 3 0 14 )
摘 要:目的 研究黄芪在体外诱导间质干细胞 (MSC)分化为神经元祥细胞的作用。方法 用Ficoll Paque液将成人骨髓细胞进行密度梯度离心和贴壁筛选分离出MSC ,体外扩增 ,流式细胞仪检测MSC表面抗原表达 ,采用含黄芪的无血清L DMEM诱导MSC分化为神经元样细胞。以免疫组化方法检测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)的表达。结果 MSC原代可获得 (6~ 8)× 10 5、第 8代可获得 (2~ 3)× 10 1 0 个细胞。流式细胞仪检测结果CD2 9、CD4 4、CD16 6表达阳性 ,CD14、CD34、CD4 5、HLA DR为阴性 ;扩增后的各代MSC表面抗原表达无显著性差异。黄芪诱导后 ,MSC形态上转变为典型的神经元样细胞 ,免疫组化显示诱导出的神构经元样细胞NSE、NF M表达阳性 ,NSE阳性细胞为 (87 2± 3 3) % ,NF M阳性细胞为 (89 1± 3 1) % ,GFAP阴性。结论 黄芪可在体外诱导成人MSC分化为神经元样细胞。Objective To investigate the differentitation of mesenchymal stem cells(MSC)into neuron-like cells with Astragale.Methods MSC were separated from bone marrow with density gradient centrifugation, wall sticking method and expanding culture in vitro.FACS was performed to detect the expression of surface antigens. MSC were induced to differentiate into neuron-like cells with L-DMEM culture medium containing Astragale in different concentrations. Neuron-specific enolase(NSE), neurofilament(NF-M), and glial fibrillary acidic protein(GFAP) were detected by immunocytochemistry.Results The count of MSC reached (6~8)×10 5 after primary culture and (2~3)×10 10 in 8th passage. These MSC were positive for CD29、CD44、CD166 and negative for CD14、CD34、CD45 and HLA-DR. There was no significant difference in expression of surface antigen between different generations of cell. The morphology of MSC change into neuron-like cells,positive NSE was(87.2±3.3) %, that of NF-M (89.1±3.1) %, but that of GFAP was.Conclusion MSC can be induced to differentiate into neuron like cells with Astragale.
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