胚胎大鼠中枢神经系统神经干细胞的分离培养与观察

目的建立神经干细胞分离、培养、分化鉴定技术,观察神经干细胞增殖、分化特点.方法利用无血清培养和细胞克隆培养技术,从胚胎大鼠海马、纹状体、脊髓等区分离神经干细胞,进行体外扩增培养、传代、贴壁分化观察.采用免疫细胞化学、透射电镜检测技术,观察鉴定神经干细胞和分化的神经细胞.结果从胚胎大鼠海马、纹状体、脊髓等区分离的细胞具有增殖能力,可进行传代培养,获得的细胞克隆中有nestin表达阳性细胞,透射电镜观察见典型的干细胞特征和不对称分裂现象.贴壁分化后可以出现MAP2、NSE、GFAP和GC表达阳性的细胞.结论用上述方法分离培养的神经干细胞具有自我更新和增殖能力,并具有向神经元、星形胶质细胞和少突胶质细胞分化的潜能.

ISOLATION, CULTURING ANDOBSERVATION OF THE NEURAL STEM CELLS FROM THE EMBRYONIC RAT CENTRAL NERVOUS SYSTEM

Objective:To establish the methods of isolation,culturing of neural stem cells and to observe the characteristics of multipotentialty and induced differentiation of neural stem cells.Methods:The advantage of serum free and clone culturing technology was performed to isolate,culture and passage neural stem cells from embryonic rat hippocampus,striatum and spinal cord.Immunocytochemistry,immunofluocytochemistry and transmission electron microscope were applied to identify the neural stem cells and their potentialty of differentiation.Results:The cell clones isolated from embryonic rat hippocampus,striatum and spinal cord possess the ability of proliferation,self reproduction and endour passage.Nestin positive cells can be found in the daughter clones.Both the mother clones and daughter clones showed typical morphological characteristics and asymmetric division of stem cells.After attachment they can differentiate into MAP 2,NSE,GFAP and GC positive cells.Conclusion:The methods to isolate and culture neural stem cells from embryonic rat hippocampus,striatum and spinal cord are established.The cultured neural stem cells possess the ability of proliferation,self reproduction and multipotentialty.