摘要
目的:建立良好的SD乳鼠神经细胞原代培养及缺血再灌注模型。方法:选用SD乳鼠大脑皮质进行细胞培养,采用机械研磨、过滤、胰酶消化、percoll(硅石胶态悬浮液)、密度梯度离心法分离纯化大脑皮质细胞后,选择含10%胎牛血清的DMEM/F12培养基进行原代培养。利用分离纯化的幼鼠大脑皮质,制作细胞爬片,利用免疫组化进行鉴定。结果:大脑皮质细胞贴壁生长,4～7d基本达到融合,结合形态学及免疫细胞化学鉴定,原代细胞培养97%为神经细胞。而后采用国际上公认的氧糖剥夺发制作缺血再灌注模型。结论:采用机械研磨、过滤、胰酶消化结合percoll:密度梯度离心法分离培养的大脑皮层神经细胞数量多,均一性生长佳,采用国际上公认的氧糖剥夺发制作缺血再灌注模型。为以后实验打下基础。
Objective：To establish a desirable method for culture of primary SD rat nerve cells and a model of cerebral ischemia/reperfusion.Methods：The cerebral cortex of young SD rat was processed to separate and purify nerves cells by mechanical trituration,trypsinization,filtration and percoll density gradient centrifugation.Then the purified cells were cultured in DMEM/F12 supplemented with 10% fetal calf serum and identified by immunocytochemistry and morphology method.Results：After 4-7days,the cultured cerebral cortex cells reached almost complete confluence.According to the results of cell morphology and immunocytochemistry analysis,97% of the cultured cells were nerve cells.Model of cerebral ischemia/reperfusion were established by oxygen sugar method which is recognized internationally.Conclusions：This culture system can yield rich and homogeneous cells,oxygen sugar method is good for establishing model of cerebral ischemia/reperfusion,which provide basis for further study.
出处
《海南医学院学报》
CAS
2011年第1期31-34,共4页
Journal of Hainan Medical University
基金
海南省自然科学基金(309062)
2010年度海南省科技厅重点计划项目(ZDXM20100047)~~
关键词
神经细胞
缺血再灌注模型
原代培养
硅石胶态悬浮液
密度梯度离心法
免疫细胞化学
Cerebral cell
Primary culture
PercolI
Denstity gradient centrifugation~ immunocy-tochemistry
Model of cerebral ischemia/reperfusionl
