成年大鼠脊髓神经干细胞的培养与鉴定

Culture and identification of neural stem cells from the spinal cord of adult rats

目的从成年大鼠脊髓中培养神经干细胞，并对其进行鉴定。方法采用悬浮培养神经干细胞技术，对成年大鼠脊髓组织进行干细胞培养，并通过自我增殖实验和免疫细胞化学技术进行鉴定。结果培养1-2周时，培养液中即出现神经干细胞克隆球。该克隆球有很强的自我增殖能力，可多次传代。免疫细胞化学技术证明该克隆球表达大量神经干细胞特征性的中间丝——巢蛋白（Nestin）。结论正常成年大鼠脊髓中含神经干细胞，在体外条件下可大量增殖。

Objective To culture and identify the neural stem cells in the spinal cord of adult rats. Metbotis Neurospheres were cultured from the spinal cord of adult rats by suspension culture system. Their stem cell nature was confirmed by self- renewal experiment and immunocytochemical technique. Results Neurospheres can be harvested from spinal cord of adult rats when cultured for I -2 weeks in vitro. They can be passaged and had strong self - renewal ability. Immunocytochemical test showed the spheres expressed the high level of nestin, a characteristic intermediate filament in neural stem cells. Conclusion The present study suggests the spheres we harvested are neurospheres. This result indicates the spinal cord of adult rats have neural stem cells, which can be enormously propagated in vitro.