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目的建立神经干细胞分离、培养和分化鉴定技术。观察神经干细胞生长、增殖特点。方法利用无血清培养,从胚胎大鼠海马、纹状体等区分离神经干细胞,进行体外扩增培养、传代观察。采用荧光免疫细胞化学检测技术,观察鉴定神经干细胞及其分化结果。结果从胚胎大鼠海马、纹状体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巢蛋白(nestin)表达阳性细胞,显微镜下观察见典型的干细胞特征。分化为3种神经细胞类型:神经元、胶质细胞、少突胶质细胞。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,通过鉴定确为神经干细胞,并经过分化鉴定确认。
Objective To establish a neural stem cell isolation, culture and differentiation of identification technology. Observation of neural stem cell growth and proliferation characteristics. Methods Using serum-free culture, neural stem cells were isolated from the hippocampus and striatum of embryonic rats, and cultured in vitro. Fluorescent immunocytochemistry was used to detect and identify neural stem cells and their differentiation results. Results The cells isolated from the hippocampus and striatum of embryonic rat had proliferative ability and could be subcultured. Among the cell clones obtained, nestin positive cells were observed, and typical stem cell characteristics were observed under microscope. Differentiation into three types of neurons: neurons, glial cells, oligodendrocytes. Conclusion The neural stem cells isolated and cultured by the above method have the ability of self-renewal and proliferation, and are identified as neural stem cells by identification and confirmed through differentiation.