贵州医科大学学报

2008, (04) 347-350

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胎鼠大脑皮层神经干细胞的分离培养、分化及鉴定
Isolation,Cultivation,Differentiation Induction and Identification of Neural Stem Cells from Embryonic Rat

胡晓霞;朱筑霞;陈刚;谭新杰;
HU Xiaoxia1,ZHU Zhuxia1,CHEN Gang1,TAN Xinjie2 (1.Department of Physiology,Guiyang Medical College,Guiyang 550004,Guizhou,China:2.Department of Carders' Wards,Guizhou Provincial People's Hospital,Guiyang 550002,Guizhou China)

摘要(Abstract):

目的:建立胎鼠大脑皮层神经干细胞体外分离、培养、分化及鉴定的方法,探讨神经干细胞的基本生物学特性。方法:取孕14 d胎鼠大脑皮层,吸管吹打机械分离制成单细胞悬液,采用无血清培养和胎牛血清诱导分化,应用免疫荧光技术鉴定神经干细胞及其分化的子代细胞。结果:从胎鼠大脑皮层分离的细胞呈神经巢蛋白(Nestin)免疫阳性并能在体外传代培养,血清诱导分化后的细胞可表达神经元、星形胶质细胞的特异性抗原微管相关蛋白2和胶质纤维酸性蛋白,诱导分化成的神经元能表达突触的特异性标志Synatophymin。结论:分离和培养的细胞能表达神经干细胞的特异性标志物Nestin,并且具有很强的增殖能力和多向分化潜能,属于中枢神经系统的神经干细胞。
Objective: To establish a method for isolation,cultivation,differentiation induction,and identification of neural stem cells(NSCs) from embryonic rat in vitro,and explore the basic biological features of these cells.Methods: The mesencephalon of rat with conceptus age of 13 days was taken out and dissociated into single cell suspension mechanically,and then cultured in serum-free medium.Fetal bovine serum was used to induce cell differentiation.The isolated cells and their progeny were observed by using immunofluorescence technique.Result: The isolated cells showed nestin positive reaction and could be cultured in vitro in series.After being induced by fetal bovine serum,microtubule-associated protein 2(MAP-2) and glial fibrillary acidic protein(GFAP) which are specific antigens in neuron and astrocyte expressed in these cells,and synatophymin,a specific signal of synapse,was also expressed in the differentiated neurons.Conclusion: The isolated and cultured cells are NSCs of central nervous system,and possess strong capability of proliferation and potency of multi-directional differentiation.

关键词(KeyWords): 干细胞;大脑皮质;细胞培养;细胞分化;胚胎;大鼠,Sprague-Dawley
stem cells;cerebral cortex;cell culture;cell differentiation;embryo;rats,sprague-dawley

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基金项目(Foundation): 贵州省科技厅基金项目(20043002);; 贵阳市科技局项目(2004第26号)

作者(Author): 胡晓霞;朱筑霞;陈刚;谭新杰;
HU Xiaoxia1,ZHU Zhuxia1,CHEN Gang1,TAN Xinjie2 (1.Department of Physiology,Guiyang Medical College,Guiyang 550004,Guizhou,China:2.Department of Carders' Wards,Guizhou Provincial People's Hospital,Guiyang 550002,Guizhou China)

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