新生大鼠海马神经元原代培养方法改进  

An Improved Method for Primary Culture of Hippocampal Neurons of New-born Rats

作　　者：李正伟[1] 陈劲草[1] 王媛[2] 张华楸[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经外科,武汉430030 [2]滨州医学院附属医院神经内科,滨州256600

出　　处：《神经损伤与功能重建》2012年第2期88-90,共3页Neural Injury and Functional Reconstruction

基　　金：湖北省科技计划自然科学基金资助项目(No.2008CDB198);国家自然科学基金(No.30900449)

摘　　要：目的:改进体外原代培养新生大鼠海马神经元的方法。方法:分离出生24h内的SD大鼠海马组织,用胰蛋白酶消化法和机械吹打法得到细胞悬液,接种24h后用无血清培养基培养神经元,每3d换液1次;倒置相差显微镜观察细胞形态,β-Tubulin与细胞核免疫荧光双染法鉴定培养神经元的纯度。结果:海马神经元生长状态好,纯度高达(94.57±1.52)%。结论:该方法培养的大鼠海马神经元活性好、纯度高,是理想的体外培养神经元的方法。Objective： To improve the method for primary culture of hippocampal neurons of new-born rats.Methods： Hippocampal regions of new-born SD rats（24 h） were harvested.Trypsin digestion and mechanical dissociation were combined to conduct mono-cell suspension.All plating media were removed from cultures and replaced with non-blood serum medium 24 hours later.Half of the culture medium was changed every three days.The morphological changes of the cultured neurons were observed under a light invert microscope.Double immunofluorescence staining of β-Tubulin and karyon were applied to assess the culture purity.Results： The cells grew well and the cell purity was demonstrated to be satisfactory [（94.57±1.52）%].Conclusion： The method is an ideal technique for culture of new-born rat hippocampal neurons with high activity and purity.

关 键 词：细胞培养 海马神经元 鉴定 免疫荧光 

分 类 号：R741[医药卫生—神经病学与精神病学] Q813.11[医药卫生—临床医学]