新生大鼠嗅鞘细胞的体外培养和纯化  

Culture and Purification of Olfactory Ensheasing Cells from Neonatal Rats

作　　者：林勇[1] 郭西良[2] 

机构地区：[1]昆明市第一人民医院神经内科,云南昆明650000 [2]界首市人民医院,安徽界首236500

出　　处：《昆明医学院学报》2009年第10期16-18,36,共4页Journal of Kunming Medical College

摘　　要：目的建立一种可行的体外培养和纯化嗅鞘细胞(OECs)的方法.方法利用显微外科技术,从新生大鼠脑中取出嗅球的嗅神经组织,去除脑膜和微血管,消化后细胞接种在含有10%的DMEM/F12(1:1)培养液中于体外进行培养,应用含血小板凝集抑制素、牛垂体提取液的DMEM/F12培养液,将这些细胞接种于p75包被的培养皿中纯化培养.用P75抗体免疫组织化学染色鉴定OECs,计算纯化度.结果OECs的外形主要以突起细长的双极和三极为主,随时间延长细胞生长排列呈现一定方向性.P75抗体免疫组化呈阳性.此方法获得的OECs纯度可达90%以上.结论本研究所建立的培养方法可行,适宜从新生大鼠中获得嗅鞘细胞.Objective To set up an available method of culturing the olfactory ensheathing cells （OECs） Methods We removed the olfactory nerve layer of olfactory bulbs from the head of neonatal rats using microsurgical techniques. The meningeal coverings and minute capillaries were pialed. The cells were cultured in vitro. The cells were seeded into DMEM/F12 （1：1） containing 10 % fetal bovine serum. Then the cells were seeded in the culture dish custodited by p75 with DMEM/F12 medium containing forskolin and pituitary extract. The purity of the OECs was evaluated according to the percentage of P75-immunostaining cells. Results Bipolar and tripolar ceils with long and slim enations were the main morphology of the OECs. Along culturing time prolongation, the OECs grew in a certain direction. P75 immunocytochemistry was positive. Purity of the OECs exceeded 90% with this culture method. Conclusion The culturing method is available and is fit for isolating ensheasing cells from neonatal rats.

关 键 词：嗅鞘细胞 培养 纯化 大鼠 

分 类 号：R332[医药卫生—人体生理学]