成人、成年狗鼻腔嗅黏膜成鞘细胞的纯化、培养与鉴定  

STUDIES ON PURIFIACATION AND IDENTIFICATION OF THE OLFACTORY ENSHEATHING CELLS FROM BOTH ADULT MAN AND CANINE

作　　者：王珂[1] 王君[1] 陈庆山[1] 孙宇[2] 潘胜发[2] 王丽[3] 于恩华[1] 周长满[1] 

机构地区：[1]北京大学医学部基础医学院解剖学系,北京100083 [2]北京大学第三附属医院骨科,北京100083 [3]北京大学第三附属医院耳鼻喉科,北京100083

出　　处：《解剖学报》2006年第1期26-29,共4页Acta Anatomica Sinica

基　　金：北京大学医学部"985"项目;北京市自然科学基金(7022018)资助

摘　　要：目的研究成人和成年狗嗅黏膜成鞘细胞(OECs)的分离与培养的方法,为探索在人体和大型动物模型的自体嗅黏膜OECs移植修复脊髓损伤奠定基础。方法采用差速贴壁方法分别从成人和成年比格狗的嗅黏膜分离出OECs,分别培养25 d,在第6 d1、0 d和25 d进行形态学观察,最后进行OECs特异标记物NGFRp75的免疫细胞化学染色,鉴定成鞘细胞。结果原代培养的成人和成年狗嗅黏膜OECs在培养10 d后明显增多,25 d的OECs成熟,具有很长的突起,其形态多为双极和三极,NGFRp75的免疫组织化学染色呈阳性。本实验条件下,狗嗅黏膜OECs的生长状态比成人的好。而未进行差速贴壁的培养细胞则几乎均呈成纤维样细胞。结论差速贴壁的方法可以分离出较为纯化的成人、成年狗鼻腔嗅黏膜OECs。Objective To investigate the technical methods for culturing and purifying the olfactory ensheathing cells （OECs） from the adult canine and human olfactory epithclium. To establish a basis for future studying the transplantation of peripheral （OECs） to repair the spinal cord injury in human. Methods Purifying the OECs from the olfactory epithelium of adult canine and man according to their different attachment time with other types of cells. Culturing for 25 days, observed at 6d, 10d and 25d, and immunostained with NGFRp75 antibody to identify the OECs. Results The number of cultured olfactory epithelium OECs from both adult canine and man were increased much more after 10 days of culture, and its sharp showed to be bi-polar or tripe-polar and are immunopositive to NGFRp75 antibody. The in vitro OECs of canine grew better than that in man＇ s in the present conditions. Conclusion The method of different attachment time seems available in purifying olfactory ensheathing cells from both the adult canine and man olfactory epithelium.

关 键 词：嗅成鞘细胞 嗅黏膜 细胞培养 免疫组织化学 人 狗 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]