人胚嗅球成鞘细胞的体外培养  

In vitro culture of human embryonic olfactory ensheathing cells

作　　者：尹国栋[1] 汤逊[1] 林月秋[1] 徐永清[1] 周田华[1] 

机构地区：[1]解放军成都军区昆明总医院全军骨科中心,云南省昆明市650032

出　　处：《中国临床康复》2006年第25期10-11,16,i0001,共4页Chinese Journal of Clinical Rehabilitation

基　　金：云南省自然科学基金资助项目(2002C0070M)~~

摘　　要：目的:评估人胚嗅球成鞘细胞的分离与培养方法。方法:试验于2005-03/2005-08在昆明总医院中心实验室完成。选取自愿捐献自然流产3～5个月的胚胎,分离原代嗅球成鞘细胞,利用延迟差速贴壁法结合阿糖胞苷抑制法对细胞进行纯化,采用免疫组织化学方法鉴定细胞并计算其P75阳性率。结果:①人胚嗅球成鞘细胞的形态学观察:原代培养5d的人胚嗅球成鞘细胞胞体呈长梭形,多极状,立体感强,折光性好。8d后,胞体变大,突起变长,并逐渐相互交织成网。14d后细胞主要表现为双极、三极形态。②人胚嗅球成鞘细胞免疫细胞化学染色及纯度鉴定:贴壁第2天,第7天的细胞染色,P75阳性达到95%以上。第14天细胞染色阳性率降至90%。25d则仅为50%。结论:应用延迟差速贴壁法与阿糖胞苷化人胚嗅鞘细胞的方法,分离培养人胚嗅球成鞘细胞是一种较为稳定可靠的方法。AIM： To investigate the in vitro isolation and culture methods of human embryonic offactory ensheathing cells. METHODS：This experiment was conducted at the Central Laboratory, Kunming General Hospital from March 2005 to August 2005. Embryos were harvested from the 3 to 5 month natural abortus, which were donated voluntarily. Primary olfactory ensheathing cells were isolated and purified with delayed differential attachment combined with cytosine arabinoside inhibiting. These cells were identified with immunohistochemical method and their P75 positive rate was calculated. RESULTS ：（1）Morphological observation of human embryonic olfactory ensheathing cells： Human embryonic olfactory ensheathing cells cultured for 5 days presented fusiform and multipolar. They had good stereoscopical feeling and refraction. Eight days later, their body became larger and the increased process wove into net each other. Fourteen days later, cells mainly presented bipolar and tripolar. （2） Immunohistochemical staining and purification identification, of human embryonic olfactory ensheathing cells： At 2 and 7 days after attachment, 95% got P75 positive. Positive rate of cellular staining decreased to 90% on day 14 and only 50% on day 25. CONCLUSION： Delayed differential attachment combined with cytosine arabinoside-treated human embryonic olfactory ensheathing cells method is stable and reliable in isolation and culture of human embryonic olfactory ensheathing cells.

关 键 词：脊髓损伤 人胚嗅球成鞘细胞 细胞培养 

分 类 号：R651.2[医药卫生—外科学]