摘要
目的:探索成人骨髓间充质干细胞(MSCs)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-1)密度梯度离心法从人的骨髓中分离出MSCs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。逐渐加入表皮生长因子(EGF)、β-巯基乙醇(BME)和全反式维甲酸(ATRA)诱导MSCs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(NSE)、神经巢蛋白(nsetin)、胶质纤维酸性蛋白(GFAP)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,MSCs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。
Aim ： To investigate the ferminal differentiation of human adult mesenchymal stem cells （hMSCs） into neuron like cells in vitro. Methods： MSCs were isolated from human adult bone marrow through density gradient centrifugation with Ficoll paque（1.077 g·mL^-1）and then amplified. Collected the second passage MSCs, were identified by the surface markers of MSCs measured by flow cytometry. MSCs were induced by adding EGF, BME and ATRA gradually and then differentiated into neuron like 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, CD 166 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 inducible rate.
出处
《中国临床神经科学》
2007年第4期410-415,共6页
Chinese Journal of Clinical Neurosciences
基金
江西省科技厅资助项目课题(编号:200401)
