新生大鼠海马和皮层神经元的原代培养及鉴定  

Primary culture method and identify in neonatal rat hippocampal and cortical neurons

作　　者：李春莉[1] 杨宝峰[2] 张景海[1] 焦军东[2] 黄文淑[1] 吴春福[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016 [2]哈尔滨医科大学药理教研室,黑龙江哈尔滨150086

出　　处：《沈阳药科大学学报》2011年第4期299-304,共6页Journal of Shenyang Pharmaceutical University

基　　金：国家自然科学基金(No.81073081);教育部博士点基金(No.20092134120001)

摘　　要：目的建立一种条件简单易存活,适用于生理、药理学研究的原代神经元培养方法。方法应用新生(48 h以内)大鼠采用急性分离、胰酶消化的方法培养海马和皮层神经细胞,并结合膜片钳及RT-PCR技术鉴定所培养的神经元电压依赖性离子通道的表达情况。结果体外培养的神经细胞生长状态良好,免疫荧光染色鉴定结果表明该方法培养的神经元纯度可达90%以上;全细胞电压依赖性离子通道表达情况良好。结论应用该方法培养神经细胞操作简单并保持了良好的生理特性,可用于神经元生理特性考察及药理实验研究,是体外培养神经元较理想的方法。Objective To introduce a simple primary culture technique suitable to neurons of neonatal rat hippocampal and cortex tissues.Methods Low concentration and prolonged time of trypsin digestion and mechanical dissociation were adopted to conduct monolayer primary culture and its expression of ion channels and electrophysiology characters were observed by using and RT-PCR the whole-cell patch clamp recording techniques.Results Under the condition in vitro,neonatal rat neurons showed similar form properties to their counterparts in vivo and typical nerve fiber net was formed.Cultivated neurons were clear and expressed different voltage-gated ion channels.Conclusion The method is simple and keeps the normal structure and function characters.This technique is an ideal method for culture in vitro for neurons.

关 键 词：神经培养 海马 皮层 鉴定 表达 

分 类 号：R96[医药卫生—药理学]