胚胎大鼠皮质神经元原代培养及神经元鉴定  

The primary culture of embryonic rat cortical neurons and neurons identification

作　　者：黄清霞[1] 卢靖[1] 张冬梅[2] 王佳雯[3] 王健[2] 

机构地区：[1]长春中医药大学 [2]长春中医药大学附属医院 [3]长春中医药大学附属医院实验中心,吉林长春130021

出　　处：《中国实验诊断学》2016年第1期1-3,共3页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金项目(81473576);吉林省科技厅自然科学基金项目(20150101251JC);吉林省卫生计生委科研计划项目(2014Z100);吉林省中医药管理局青年基金项目(2014-Q2)

摘　　要：目的对原有的胚胎大鼠神经细胞原代培养进行改进,建立一种高效、稳定的的原代培养方法。方法自精确控制胎龄的大鼠胚胎大脑中分离出皮质,采用胰酶二次消化法制备神经细胞悬液。并选用神经元特异性烯醇化酶免疫荧光化学法对神经细胞进行鉴定。结果与单次消化法分离细胞及运用DMEM+10%胎牛血清重悬细胞进行种板的原方法相比,方法改进以后,细胞提取数量明显增加,且纯度较高。种板后细胞形态正常且能长期存活。结论该方法结果理想,简易高效,值得借鉴和推广。Objective To establish a highly efficient and stable method for the primary culture by modified the origi- nal method of cultureing embryonic rat cortical neurons. Methods Cerebral cortex isolated from the precise control of gestational age was dissociated to single cells by the twice-enzyme isolation method. Immunofluoreseence of neuronspe cific enolase chemical methods was used to identify neurons. Results The impro-~ed method could markably increase the number of cells and the purity compared to the original method of single digestion method and suspending cells use the DMEM＋10~fetal bovine serum. The plateing neurons was eumorphism and has long-time livability. Conclusion The experimental method has a ideal result also is simple and efficient that is worth to reference and generalize.

关 键 词：胎鼠 神经元 原代培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] Q813.11[医药卫生—基础医学]