大鼠神经干细胞的分离培养和分化鉴定

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。

Isolation,incubation,differentiation and identification of neural stem cells in rats

Objective To establish the methods of isolation,culturing,differentiation and evaluation of neural stem cell,observe the characteristics of growth and multiplication of neural stem cells.Methods The serum-free culturing technology was performed to isolate stem cells from embryonic rat hippocampus,striatum and culture them in vitro. Fluorescence immunocytochemistry was applied to identify the neural stem cells and the produce of differentiation.Results The cell clones isolated from embryonic rat hippocampus,striatum possessed the ability of proliferation,selfreproduction.Nestin-positive cells could be found. Through microscopy the cell clones showed typical characteristics of neural stem cells. Produce of differentiation:nerve cell,glial cell and oligodendrocyte.Conclusion The method is able to isolate and culture neural stem cells from embryonic rat hippocamus and striatum,the cells possess the ability of proliferation,selfreproduction. being identified as neural stem cells.