胎鼠大脑皮质神经元的体外培养及鉴定

为了建立SD大鼠大脑皮质神经元的分离、培养及免疫细胞化学鉴定技术,本研究通过分离孕15 d SD胎鼠大脑皮质组织,经机械吹打后,加入含有5％马血清、5％小牛血清的MEM培养基接种在24孔培养板中进行原代培养,并于第3 d在培养基中加入阿糖胞苷抑制胶质细胞的生长,于第9 d将细胞固定后采用免疫细胞化学技术检测神经元特异性烯醇化酶(NSE)的表达。结果表明从孕15 d SD胎鼠大脑皮质分离的神经元纯度高,生长旺盛,形态典型,90％以上表达NSE免疫阳性。该结果提示我们建立的培养大鼠大脑皮质神经元的方法易于标准化操作,结果稳定,值得推广应用。

ISOLATION AND IDENTIFICATION OF NEURONS FROM THE CEREBRAL CORTEX OF EMBRYONIC RATS

In order to establish a method of isolating, culturing and identifying neurons from the cerebral cortex of E15 rats,cells were incubated in a 24-well-plate from the cerebral cortex of E15 rats by using a MEM mediumadded with 5% equineserum, 5% bovine serum in the present study. Cytosine arabinoside was added to the medium on day 3 during culture to restrainthe growth of glia cells. The cultured cells were then fixed on day 9 and immunocytochemically stained with neuronal specificenolase (NSE) antibody. The present results indicate that above 90% of the cells isolated from the cerebral cortex of E15 ratsgrew well and expressed NSE immunoreactivity. Therefore, it implies that a novel and practical method to culture pure neuronsfrom E15 rats in our study is constructed, and the results are stable and worth of application.