摘要
目的 优化海马神经元原代培养方法。方法 取孕17～18d胎鼠海马，经过剪碎、消化、离心和吹打过滤后种板，根据孵育后更换为无血清培养基的时间不同分为A组和B组。A组4h后更换，B组12h后更换。观察并记录第2、3、4、5天两组神经元生长状况，并于第7d用神经元特异性烯醇化酶荧光染色法鉴定神经元，计算其纯度。结果 （1）4～6h后细胞已贴壁生长，2d后胞体增大，3d时突起长出，5d后突起变长变粗并有分支形成，相互连接呈网状。（2）7d后行免疫荧光染色，为阳性结果，证实为神经元，且A组神经元纯度显著高于B组（P＜0.05）。结论 改进后的方法可培养出生长状况良好且纯度更高的神经元。
Objective To optimize the primary culture method for hippocampal neurons. Methods Hippocampus was harvested from fetal rats. They were obtained by enzymatic digestion and divided into groups A and B according to different times for replacing serum-free medium （4h after seeding in group A and 12h in group B）. The cell growth was recorded on the 2nd, 3rd, 4th, and 5th days. Fluorescence assay was used to identify the cells with neuron specific enolization and determine the purity on the 7th day. Results After 4？6h, the cells attached to the wall. The cell body was enlarged on 2nd day. The nervous process erupted on 3rd day and extended into reticulation on 5th day. The fluorescence assay identified the neurons and indicated that the purity of group A was obviously higher than group B （P〈0.05）. Conclusion The improved technique can cultivate well-growth hippocampal neurons with higher purity.
出处
《中华老年多器官疾病杂志》
2016年第1期52-55,共4页
Chinese Journal of Multiple Organ Diseases in the Elderly
基金
2014年度上海市卫生和计划生育委员会科研课题重点项目（201440022）
关键词
神经元
培养
胎鼠
neurons
culture
fetal rats
