成年大鼠嗅球嗅鞘细胞的分离、培养、纯化与生物学特性的研究  

Isolation,Culture,Purification and Biological Characteristics of Olfactory Ensheathing Cells from the Adult Rat Olfactory Bulbs

作　　者：范志海[1] 沈忆新[1] 陆政峰[1] 成茂华[1] 董启榕[1] 

机构地区：[1]苏州大学附属第二医院骨科,江苏苏州215004

出　　处：《苏州大学学报（医学版）》2007年第4期547-551,共5页Suzhou University Journal of Medical Science

摘　　要：目的探讨成年大鼠嗅球嗅鞘细胞(OECs)的分离、培养和纯化方法,并观察其形态学变化及免疫细胞化学特性。方法采用原代培养技术,从成年大鼠的嗅球分离培养OECs,用差速贴壁+Thy1.1抗体及补体等方法纯化培养OECs,并用碱性成纤维细胞生长因子(bFGF)扩增OECs,倒置相差显微镜下观察OECs的形态变化特点及生长情况,采用GFAP、NGFRp75、S100等抗体行免疫细胞化学染色,根据NGFRp75免疫细胞化学染色结果统计培养的OECs的纯度,并计算OECs的增殖率。结果体外培养的成年大鼠OECs主要为双极或三极细胞,其突起细长。差速贴壁+Thy1.1抗体及补体纯化法的纯化效率高于其他两种。免疫细胞化学染色显示,培养的成年大鼠OECs呈现GFAP、NGFRp75、S100蛋白染色阳性。bFGF能明显促进OECs的增殖(P<0.01),对OECs的形态、纯度没有明显影响。结论差速贴壁+Thy1.1抗体及补体纯化法是一种高效率的纯化培养OECs的方法,bFGF能明显促进OECs的增殖。Objective To culture and purify the olfactory ensheathing cells （OECs） from the adult rat olfactory bulbs in vitro, and to investigate the morphological changes and the immunocytochemical properties of the cultured OECs. Methods The OECs were dissociated from the adult rat olfac- tory bulbs. The primarily cultured OECs were purified based on the differing rates of attachment of the various harvested cell types and （or） anti-Thy 1.1-mediated complement lysis. The morphological changes of the cultured OECs were observed under a phase contrast microscope at different culture time. The OECs were identified by GFAP, NGFRp75 and S100 immunocytochemistry. The purity of the OECs was evaluated according to the percentage of NGFRp75 immunostaining cells. Results The OECs displayed a very characteristic morphological appearance. Most of OECs were bipolar or tripolar with long and thin processes. The OECs puritification by the method based on the differing rates of attachment of the various harvested cell types with anti-Thy 1.1-mediated complement lysis was more efficient than the others, bFGF could facilitate OECs growth, and could not affect the proportion of positive of OECs. Conclusion The method of purification for OECs through combining with the differing rates of attachment of the various harvested cell types with anti-Thy 1.1-mediated complement lysis is effective for the enrichment of OECs. bFGF can facilitate OECs growth.

关 键 词：嗅鞘细胞 嗅球 细胞培养 生物学特性 

分 类 号：R329.34[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]