胚胎大鼠背根神经节神经元分离、纯化与培养  

The dissection,purification and culture of dorsal root ganglion neurons from embryonic rats

作　　者：王士杰[1,2] 黄丽辉[1,2] 雷雳[1] 王鸿[1,2] 范尔钟[1,2] 李颖[1,2] 

机构地区：[1]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科耳鼻咽喉头颈科学教育部重点实验室(首都医科大学),北京100730 [2]北京市耳鼻咽喉科研究所,北京100005

出　　处：《中国耳鼻咽喉头颈外科》2010年第12期656-658,共3页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：国家自然科学基金项目(30772406);国家"十五"科技攻关项目(2004BA720A18-02);国家"十一五"科技支撑计划(2007BAI18B12)联合资助

摘　　要：目的建立一种简单、稳定、高效的胚胎大鼠背根神经节神经元原代培养方法。方法摘取14～16 d的胚胎大鼠背根神经节,采用0.25%胰蛋白酶加0.4%胶原酶II消化,制成单细胞悬液,接种于Neuralbasal培养基中,24h加入有丝分裂抑制剂阿糖胞苷纯化细胞,应用神经元特异性烯醇化酶(neuronal specific enolase,NSE)进行免疫细胞化学染色,鉴定细胞纯度。结果体外培养的背根神经节神经元生长状态良好,可存活6周,纯度可达到(91.12±0.23)%。结论本实验方法简单、稳定、高效,可以获得高纯度的背根神经节神经元培养物。OBJECTIVE To establish an simple、 efficient、reliable method for the purification culture system of dorsal root ganglion neurons derived from embryonic rats.METHODS Dorsal root ganglions harvested from embroynic rats were digested with the mixture of trypsin and collegonease II,then turned into single cell suspension and plated in neuralbasal media.Cultured dorsal root ganglion neurons were purified by cytosine arabinoside for 24 hours.The purificational rate was evaluated according to cell count and neuronal specific enolase（NSE） immunocytochemistry stain.RESULTS Cultured dorsal root ganglion cells could survive healthily.The purification rate of neurons was（91.12±0.23）%.CONCLUSION The method,which is used for cultue and purification of DRGn,is a simple、efficient、reliable way.Using it could obtain highly purifacational neurons.

关 键 词：动物实验 神经节 脊 细胞培养 磷酸丙酮 酸水合酶 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]