新生大鼠神经干细胞的分离培养与观察

Isolation, culture and observation of the neural stem cells from the central nervous system of neonatal rat

目的 建立神经干细胞的分离、培养及分化鉴定技术 ;观察神经干细胞的生长、增殖及分化的特点。方法 分离新生SD大鼠脑室下区的组织 ,经消化及机械吹打后 ,采用原代贴壁及传代悬浮的方法 ,培养获得细胞克隆。应用免疫细胞化学方法及透射电镜检测技术 ,鉴定神经干细胞和分化的神经细胞。结果 从新生SD大鼠脑室下区分离的组织 ,经原代和传代培养均可形成细胞克隆 ,并具有增殖的能力。原代和传代细胞抗原 nestin呈阳性。分化后的细胞可表达神经元、星形角质细胞和少突角质细胞的特异性抗原。结论 用此方法分离的细胞具有自我更新和分化潜能 ,有很强的增殖能力 。

Aim To establish the methods of isolation and culture of neural stem cells and to observe the characteristics of growth proliferation, and induced differentiation of neural stem cells.Methods The neonatal rat subventricular zone tissues were dissociated mechanically and enzymatically. The primary adhesion and passage suspension culture technique were performed to acquire cell clones. Immunocytochemical staining and transmission electron microscope were used to identify the neural stem cells and their differentiated offspring.Results Cell clones could from the neonatal rat subventricular zone tissue after primary and passage culture. There clones were nestin positive and possessed the ability of proliferation and self renewal. The differentiatied cells expressed neurons, astrocytes, oligodendrocytes specific antigen . Conclusion The neural stem cells isolated and cultured by using the present methods have the ability of self renewal and differentiation.