大鼠脑室下区神经干细胞的分离培养与分化

Isolation, Culture and Differentiation of the Neural Stem Cells from the Central Nerve System of Neonatal Rat

目的:从新生SD大鼠脑室下区分离并培养神经干细胞,观察其生长、增殖及分化.方法:采取无血清培养和单细胞克隆技术,采用原代贴壁及传代悬浮方法,培养获得细胞克隆.利用免疫细胞化学方法鉴定神经干细胞.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,且具有增殖能力.原代和传代细胞巢蛋白(Nestin)、神经元特异性烯醇化酶(Neuronspecificenolase,NSE)抗原呈阳性.神经干细胞可分化为神经元.结论:用上述方法分离的细胞具有自我更新能力和分化潜能以及很强的增殖能力,属于中枢神经系统的干细胞.

Aim: The neonatal rat sub ventricular zone tissues were dissociated mechanically and systematically and to observe the characteristics of growth proliferation and induce differentiation of neural stem cells. Methods: To establish the methods of serum -free incubation and single-cell cloning technique, isolation and culture of neural of stem cells to acquire cell clones. Immunity technical 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 sub ventricular zone tissue after primary and passage culture. There clones were nesting-positive and possessed the ability of proliferation and self renewal. The differentiated cells expressed NSE specific antigen. Conclusion: The neural stem cells isolated and cultured by using the present methods have the ability of self-renewal and differentiation.