胎鼠海马神经元培养及其鉴定  

Identification and Culture of Hippocampus Neurons in Fetal Rats

作　　者：李玉[1] 王波[1] 但齐琴[2] 

机构地区：[1]昆明医学院第一附属医院神经外科,云南昆明650031 [2]昆明医学院神经科学研究所,云南昆明650031

出　　处：《昆明医学院学报》2011年第5期17-19,共3页Journal of Kunming Medical College

摘　　要：目的建立胎鼠海马神经元体外培养方法,观察细胞生长情况,免疫组织化学染色鉴定神经元特异性.方法取胎鼠海马,分裂获得细胞悬液,接种于24孔板,经2 h差速帖壁去除成纤维细胞,将细胞液转移并继续培养,以获娶纯度较高的海马神经元.培养第5、11、15天观察细胞和突起生长情况,用NeuN抗体以免疫组化SP二步法染色鉴立神经细胞.结果培养头3 d.细胞生长缓慢,5 d时细胞开始生长,可见突起,11 d时突起连成网状.经Neun染色培养细胞呈阳性反应,证明是神经元.结论本方法获取生长良好,纯度较高的海马神经元.Objective To establish the protocol for culturing hippocampus neurons in vitro and explore the neuronal growth character istics.Methods Tissues from hippocampus of fatal rats were harvested,then digested into cell suspension by using 0.125% trypsin.After planted into wells for 2 hours,cell suspension was transferred into next well for continuing incubation.The cell growth was observed at day 5,11 and 15.Neurons specific Enolase（NSE）antibody,recognized specifically NSE antigen in cultured cells,was used to identify neurons by SP-two-step staining method.Results After incubated for 2 hours,transferred suspensions in wells showed some pure neurons,which is identified by NSE staining.However,they grew slowly during 1～3 days,then grew well from 5th day,with gradual increasing the neuritis outgrowth.Conclusion Purified neurons with neurite outgrowth states can be acquired by this method.

关 键 词：胎鼠 海马神经元 NEUN 培养 

分 类 号：Q831.11[生物学]