成年大鼠嗅神经鞘细胞纯化培养的研究  

A STUDY ON THE PURIFICATION OF ADULT RAT OLFACTORY ENSHEATHING CELLS IN VITRO

作　　者：杨浩[1] 鞠冬阳[1] 李静雯[1] 程希平[1] 游思维[1] 鞠躬[1] 

机构地区：[1]第四军医大学全军神经科学研究所,西安710032

出　　处：《神经解剖学杂志》2005年第2期180-184,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(No. 30100195; 30371466)资助项目

摘　　要：嗅神经鞘细胞(olfactoryensheathingcells,OECs)是目前用于神经再生研究较为理想的移植细胞,为了建立一种可以有效获得高纯度、高均一性的OECs培养方法,本实验采用2. 5月的大鼠嗅球为实验材料进行OECs培养,同时结合OECs的生物学特点在培养8d后,通过用硝酸纤维素膜吸附p75抗体,再对培养的OECs进行免疫亲和吸附和对成纤维细胞进行补体杀伤来纯化OECs, 在纯化后分别对培养2、4、6、8d的OECs进行p75免疫组化染色和纯度鉴定。实验结果发现,本研究采用的OECs纯化培养方法所获得OECs纯度在4个不同培养时间点都达到98%以上,说明此方法是一种高效率的纯化培养OECs的方法。Olfactory ensheathing cells (OECs) are nowadays an optimal candidate for transplantation into CNS injury in nerve regeneration. The present study is aimed to establish an effective culture method to acquire highly and uniformly enriched population of OECs for study on cell transplantation. Primarily cultured OECs were dissociated from olfactory bulb(OB) of the 2.5 month old rats. After cultured 8 days in vitro , the OECs were purified away from other cell types present in the cultures according to its biological characteristics as follows. The nitrocellulose membrane was coated with p75 antibody , OECs purification was used to eliminate much of contaminating cells by immunoadsorption and subsequent complement killing methods. Then Purity of OECs was identified with immunocytochemical staining after purified OECs were cultured 2,4,6 and 8 days in vitro , respectively. Our results show that the OECs purity reach above 98% at 4 distinct culture time points, respectively by using the modified method. This method is highly effective for enrichment of OECs.

关 键 词：成年大鼠 嗅神经鞘细胞 纯化培养 细胞培养 中枢神经系统损伤 神经移植 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]