【作者】 杨志刚；

【导师】 吴晓牧；

【作者基本信息】 南昌大学 ， 神经病学， 2006， 硕士

【摘要】 目的：探索成人骨髓间充质干细胞的分离培养及诱导分化为神经元样细胞的体外条件。 方法：采用Ficoll淋巴细胞分离液(1.077g／ml)密度梯度离心法从人的骨髓中分离出骨髓间充质干细胞进行体外扩增。收集第2代细胞流式细胞仪检测MSCs表面标志。逐渐加入EGF、BME和ATRA诱导骨髓间充质干细胞向神经元细胞分化，通过免疫细胞化学法鉴定诱导后细胞神经元烯醇化酶、神经巢蛋白、胶质纤维酸性蛋白的表达情况。 结果：流式细胞仪检测结果显示培养后的MSCs强表达CD29；较强表达CD44、CD166和CD105；不表达CD45、CD34和HLA-DR。诱导剂诱导后，骨髓间充质干细胞分化为具有典型的神经元形态的细胞，免疫细胞化学显示78％的细胞NSE表达阳性；大约51％细胞Nsetin表达阳性；所有细胞GFAP均阴性表达。 结论：成人MSCs在体外可定向诱导分化为神经元样细胞，且具有较高的阳性分化率。更多还原

【Abstract】 Objective: To investigate the isolation and culture of human adult mesenchymal stem cells (hMSCs) and then differentiate it into neuron like cells in vitro.Methods: MSCs were isolated from human adult bone marrow through density gradient with Ficoll paque (1.077g/ml) and then amplified . Collected the second passage cells, identified surfacemarkers of MSCs by flow cytometry. Added EGF、BME and ATRA gradually and then differentiate it into neuron cells. The expressions of neuron specifice nolase (NSE)、nestin and glialfibriliary acidic protein (GFAP) were detected by immunocytochemistry.Results: By flow cytometry analysis, MSCs were strongly positive expressed of CD29, positive for CD44、CD166 and CD105 and negative for CD45、CD34 and HLA-DR. After treatment with EGF, BME and ATRA, MSCs could be differentiated into cells with typical neuronal morphology, 78% of the cells expressed NSE, 51% of the cells expressed Nsetin, none of them expressed GFAP.Conclusion: MSCs can be induced to differentiate into neuron like cells with high positive rate of induced differentiation.更多还原