豚鼠鼻腔上皮嗅神经鞘细胞的体外培养及鉴定  

Olfactory Ensheathing Cell From Nosal Cavity of Guinea Pig Culture and Appraisal in Vitro Study

作　　者：朱勇[1] 王琳[1] 

机构地区：[1]西安医学院,陕西西安710021

出　　处：《延安大学学报（医学科学版）》2008年第2期6-7,11,共3页Journal of Yan'an University:Medical Science Edition

基　　金：陕西省卫生厅科研基金2006年资助课题(06D14)

摘　　要：目的探讨成年豚鼠嗅球嗅神经鞘细胞(Olfactory ensheathing cells,OECs)取材、体外培养及纯化方法,为以豚鼠作为实验模型的神经移植实验及进一步研究其生物学特性打下基础。方法在解剖显微镜下仔细分离出嗅球的第一二层,经剪碎、消化及吹打后去除沉淀,并行培养前差速贴壁法初步纯化及培养后第7-10d用消化法行进一步纯化,用神经生长因子受体蛋白(p75NGFR)行免疫组织化学鉴定及估算OECs纯度。结果体外培养成年豚鼠OECs形态与成年大鼠基本相似;未经纯化处理的嗅球组织中的OECs,在培养2周时被大量增殖的成纤维细胞所覆盖和取代。而经过精细取材、培养前纯化和培养后纯化的综合处理,可使OECs纯度达到70%。结论精细取材和综合纯化可培养出纯度较高的OECs,为以豚鼠作为实验模型的神经移植实验及进一步研究其生物学特性打下基础。Objective To explore the methods of dissociation,culture in vitro and purification of olfactory ensheathing cells(OECs) from adult guinea pig olfactory bulbs so as to lay foundations of further research on the biological characteristics of OECs!and experiment of neural regeneration based on experimental model of adult guinea pig.Methods The tissue in the first two layers of olfactory bulb was dissociated carefully under anatomic microscope,cut up and then incubated in 3mL/L collagenase I for 30 min.After centrifuged,the cell pellet was dispersed and the deposition was removed;the supernated fluid was initially purified with differential attachment method before culture,and the further purification was made with digestive method from the 7th to 10th day after culture in vitro;and the purity of OECs was estimated and appraised by immunostaining positively for NGF receptor(p75NGFR).Results The morphological features of OECs in vitro from adult guinea pig olfactory bulbs were basically similar to that of the cells from adult rats;and the OECs without purified were mostly covered and replaced by proliferated fibroblasts.While through refined dissociation and synthetic purification both before and after culture,the purity of OECs reached about 70%.Conclusion Through refined dissociation and synthetic purification,a higher purity of OECs from adult guinea pig olfactory bulbs can be obtained,which lays foundations of further research on the biological characteristics of OECs and experiment of neural regeneration based on experimental model of adult guinea pig.

关 键 词：嗅神经鞘细胞 豚鼠 细胞培养 免疫组织化学 

分 类 号：R745[医药卫生—神经病学与精神病学]