摘要
目的 研究体外原代培养新生大鼠海马神经元的方法，并观察其发育分化过程中形态学的变化。方法 取出生24h内的Wistar大鼠分离海马，消化后差速贴壁，种植在涂有多聚-L-赖氨酸的盖玻片上，于不同时间在倒置相差显微镜下观察形态变化；采用NSE免疫细胞化学技术鉴定神经元。结果 神经元12～24h后大部分贴壁，随时间延长形态多变，突起逐渐增多，神经元突起间互相接触形成网络。培养第7～10天时细胞最为丰满，至28天时培养液中有悬浮的细胞碎片。NSE鉴定结果显示神经元纯度为（92．3±6．8）％。结论 该方法简单易行，神经元纯度较高，可作为神经元体外培养的良好模型用于以后的研究。
Objective To establish a primary culture method of hippocampal neurons of new-born rats, and to observe the morphological characteristics at different developmental and differential stages. Methods The hippocampus was digested and the cells were seeded in a flask. The neurons were then transferred and re-seeded on cover coated with poly-L-lysine. The neurons were identified by lase （NSE）. The morphology was observed under phase-contrast polyclonal antibody against neuron specific enomicroscope. Results A large number of hippoeampal neurons began to adhere to the cover glasses after 12 - 24 hours. They showed different shapes-shuttle, trilength to the plate. Their processes connected into nets and different in and thickness. They were well developed on the 7th - 10th day and survived as long as 28 days after seeding. Immunoeytochemistry of NSE proved high purity. Conclusion The culture method of new-born rat hippoeampal neurons in vitro is successful and can be used as an in vitro model of research.
出处
《基础医学与临床》
CSCD
北大核心
2007年第3期329-332,共4页
Basic and Clinical Medicine
