大鼠海马神经元的原代培养及细胞鉴定  

Primary culture method for hippocampal neurons of rats and identification of cells

作　　者：王翀[1] 赵康峰[1] 顾雯[1] 李玲[1] 张宏伟[1] 白雪涛[1] 

机构地区：[1]中国疾病预防控制中心环境与健康相关产品安全所,北京100021

出　　处：《环境与健康杂志》2013年第9期767-769,F0003,共4页Journal of Environment and Health

基　　金：国家自然科学基金(81072260)

摘　　要：目的建立一种大鼠海马神经元的分离及原代培养方法，并进行细胞鉴定来确定其特质性，为进行海马神经元的体外研究提供技术支持。方法选用出生24h内的Wistar大鼠，分离双侧海马，经胰酶消化后用含10％马血清的DMEM／F12（1：1）培养基培养，12h内换成无血清培养基培养48h，加入阿糖胞苷处理24h，第5-8天用免疫组织化学法鉴定微管相关蛋白-2（MAP-2）的表达。结果接种4h后细胞开始贴壁，有的长出细小突起，胶质细胞生长活跃，换为无血清培养及阿糖胞苷处理后胶质细胞生长抑制，海马神经元生长占优势，随着培养时间的增加，细胞突起逐渐增长及网状联络密集。培养至3周细胞逐渐死亡。MAP-2鉴定表达阳性。结论出生24h内的乳鼠原代海马神经元的分离及培养方法可靠，可作为神经元体外培养的良好模型，为体外研究提供可靠的技术支持。Objective To establish a method of seperation and primary culture for hippocampal neurons of rats in vitro and identify them and provide technique supports for hippocampal neurons study in vitro. Methods Hippocampal neurons of 1-day-old Wistar rats were collected and cultured with DMEM/F12（I：I） with 10% horse serum, and within 12 h followed by serum-free medium for 48 h, then treated with cytosine arabinoside for 24 h, microvascular protein-2 （MAP-2） was identified with immunohistoehemistry method on day 5-8. Results Adherence was seen after 4 hours of culture and some neurons had small enations, glial ceils grew actively and then were inhibited by serum-free medium and cytosine arabinoside, while hippocampal neurons grew well instead. The axon/dendrite developed well and connected with each other. The neurons were dead gradually three weeks later. MAP-2 expressed in hippocampal neurons in this experiment. Conclusion The method of seperation and primary culture of hippocampal neurons is reliable and can be used as good model for neurons culture in vitro.

关 键 词：海马神经元 原代培养 微管相关蛋白-2 细胞鉴定 

分 类 号：R994.6[医药卫生—毒理学]