大鼠皮层神经元和胶质细胞混合培养牵张损伤模型的建立  

The establishment of an in vitro model of stretch-induced injury on cultured cortical neurons and astrocytes in rats

作　　者：范东东[1] 钟春龙[1] 夏蓉[1] 张明坤[1] 崔振文[1] 江基尧[1] 罗其中[1] 

机构地区：[1]上海交通大学医学院附属仁济医院东院神经外科,上海200127

出　　处：《立体定向和功能性神经外科杂志》2010年第4期214-217,共4页Chinese Journal of Stereotactic and Functional Neurosurgery

基　　金：国家自然科学基金项目(编号:30500525;30670719);上海市科委基础重点研究计划项目(编号:07JC14038);上海市青年科技启明星计划项目(编号:05QMX1428)

摘　　要：目的建立大鼠皮层神经元胶质细胞混合培养牵张损伤模型。方法提取出生时间24h到48h的SD大鼠大脑皮层组织行神经元和胶质细胞原代混合培养,根据Ellis方法采用CICⅡ型细胞损伤装置建立大鼠皮层神经元和胶质细胞混合培养牵张损伤模型,根据硅胶膜的变形距离将实验组分为轻度组、中度组和重度组。同时设立对照组,不作牵张损伤。分别在不同时间进行神经细胞的PrI染色和测定培养上清液中的LDH含量,以确定损伤的程度。结果随着牵张损伤程度加重,PrI红染细胞数逐渐增多,同时细胞培养液中LDH释放量也逐渐增多。结论成功建立了大鼠皮层神经元胶质细胞体外牵张损伤模型,可分别模拟轻、中、重度损伤。使用方便,可重复性好,可用于颅脑创伤的病理机制和药物作用的进一步研究。Objective To establish an in vitro model of stretch-induced injury on cultured cortical neurons and astrocytes in rats.Methods Rat cortical neurons and astrocytes from 1 to 2 days old rats were cultured,then were plated in Bioflex plates with silastic bottom after purification.Then the injury group was divided into mild group、moderate group and severe group.The control group did not take injury.Extent of injury was qualitatively assessed by Propidium Iodide（PrI） staining and lactate dehydrogenase（LDH） enzyme release.Results Different degree of rapid deformation of silastic membrane and different degree of cell injury were caused by different gas pressure.With the increase of the gas pressure,PrI staining and the value of LDH release also increased.Conclusion An in vitro model of stretch-induced injury on cultured cortical neurons and astrocytes in rats has been established,and it can be used to study the pathology mechanism and drug effects after injury.

关 键 词：创伤性脑损伤 体外模型 神经元 胶质细胞 牵张损伤 

分 类 号：R651[医药卫生—外科学]