E18胚胎大鼠皮层区离体神经元培养  

Culture of Cortical Neurons from E18 Prenatal Rats

作　　者：王金鹏 张勇 WANG Jinpeng;ZHANG Yong(Department of Neurobiology,School of Basic Medical Sciences and Neuroscience Research Institute,Key Lab for Neuroscience,Ministry of Education of China,National Health Commission,Peking University,Beijing 100083,China)

机构地区：[1]北京大学基础医学院神经生物学系北京大学神经科学研究所教育部神经科学重点实验室国家卫生健康委员会神经科学重点实验室

出　　处：《中国细胞生物学学报》2019年第9期1772-1778,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31771125)资助的课题~~

摘　　要：神经元细胞体外培养,尤其来自中枢神经系统内的细胞,是研究神经退行性疾病发病机制、神经再生过程以及基因工程小鼠模型等重要的实验手段。E18胚胎大鼠皮层神经元体外培养技术,主要包括前期圆玻片处理、皮层分离、细胞消化、铺种以及更换培养基等。结果发现,该研究神经元细胞生长良好,可维持生长至少20天。在神经元形态方面,树突具有较多分支,轴突在不同细胞间形成连接。通过免疫染色和共聚焦显微镜成像,该研究可观察到树突棘结构。此外,对培养的神经元进行转染实验发现,转染后细胞状态良好,转染效率在10%左右。综上,神经元体外培养技术方法可以较好地培养神经元并能维持神经元正常发育生长。体外培养的神经元细胞可用于免疫组化、基因编辑以及实时成像研究。Primary neuronal culture from prenatal rats is widely used to uncover cellular mechanisms underlying various processes in neurons such as cellular trafficking,cellular structure and protein localization.However,unambiguous results from neuronal culture depend on the health,purity and complexity of the neurons.Here we provide a protocol for isolating and culturing cortical neurons from E18 embryonic rats.We discuss detailed culture techniques including glass coverslips treatment,cortex dissection,digestion,plating and medium replacement.Under our protocol,the cultured cortical neurons could be maintained in healthy condition(neuronal morphology,extensive axonal and dendritic arbors development)for at least 20 days.The cultured neurons were tested for immunostaining and confocal imaging,we can also achieve a 10%transfection efficiency with the neuronal culture.Overall,this optimized cell culture technique can be used to culture and maintain the normal development of neurons for applications such as immunohistochemistry,gene editing as well as live imaging studies.

关 键 词：神经元 细胞培养 皮层 大鼠胚胎 

分 类 号：R74[医药卫生—神经病学与精神病学]