成人鼻腔嗅区黏膜细胞的分离培养与免疫学特点  

Isolationculture and characteristics of human olfactory mucosa cells

作　　者：王洪美[1,2] 任玉水[1] 田国忠[1] 姜晓荣[1] 曹敬丽[1] 刘婧媛[1] 周长满[3] 黄红云[1,2] 

机构地区：[1]北京市虹天济神经科学研究院临床细胞中心,北京100144 [2]北京康复中心神经外科,北京100144 [3]北京大学医学部人体解剖与组织胚胎学系,北京100191

出　　处：《解剖学报》2010年第3期374-378,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30971527)

摘　　要：目的探讨成人鼻腔嗅区黏膜细胞的分离培养方法及嗅鞘细胞(OECs)的免疫学特点。方法通过手术取成人嗅区黏膜7例,将其消化为单个细胞后采用差速贴壁法除去成纤维细胞。倒置显微镜下行形态学观察,并利用p75、S100、GFAP抗体对嗅鞘细胞进行免疫荧光化学染色,共焦显微镜下进行各种抗体阳性率的统计。结果共焦显微镜下可见嗅黏膜细胞中S100和p75阳性细胞多见,GFAP阳性细胞在嗅黏膜细胞中较少见。一般在同一细胞上可见S100和p75共同表达,而GFAP与S100、p75未见有共同表达现象;p75阳性细胞胞体大小和形态也大不相同。经统计S100和p75阳性细胞的平均百分率为(35.0±8.3)%。结论成功进行了成人鼻腔嗅区黏膜细胞的分离培养。Objective To establish a technical method to isolate and culture human olfactory mucosa cells in vitro, and to study the immunological characteristics of olfactory ensheathing cells（OECs）. Methods In this study, seven adult olfactory mucosae were obtained from nasal cavity surgery. Cells were acquired by dissection of olfactory mucosa and purified according to the different attachment time among the various cell types. The cells were observed for morphological characteristics under inverted microscope and identified using immunofluorescence staining with p75, S100 and glial fibrillary acidic protein （GFAP）. The characteristics of olfactory ensheathing cells was observed under confocal microscope. Results S100 and p75 positive cells were more common in the olfactory mucosa cells under confocal microscope, GFAP positive cells were relatively rare. In general, S100 and p75 were co-expressed in the same cells, but not the GFAP and S100, p75 expressions. The p75 positive cells presented two completely different morphology. The percentage of S100 and p75 double labeled cells was （35.0 ± 8.3） % under confocal microscope. Conclusion The olfactory ensheathing cells can be isolated from human olfactory mucosa, they are accessible sources for autologous grafting to repair nervous system disease.

关 键 词：嗅区黏膜 嗅鞘细胞 细胞培养 P75 S100 免疫荧光化学 细胞移植 人 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]