人嗅粘膜嗅鞘细胞的分离、培养及生物学特性

目的细胞移植是目前的研究热点,本文主要研究人嗅粘膜嗅鞘细胞(OM-OECS)的分离、培养、纯化、鉴定,来观察其生长情况,分析其生物学特性,为具有神经功能障碍的患者实行同种细胞移植提供一种可行的方法。方法选取意外交通事故或意外疾病、事故急性死亡3h内及经蝶窦手术的患者,征得家属同意,取鼻中隔后1/3鼻粘膜,免疫吸附法结合采用P75抗体包被的培养皿进行纯化人OM-OECs,免疫组化方法进行细胞的鉴定。结果 OM-OECs体外培养过程中,采用免疫吸附法使抗体与间充质来源的成纤维细胞进行特异结合从而将其去除,P75抗体包被的培养皿促进了OECs的贴壁使其与杂质细胞进一步分离,在体外培养10d后,细胞增殖最快,经P75、GFAP免疫组化双标染色显示呈双阳性,计数阳性细胞率达90%以上,说明从人嗅粘膜分离培养OECs具有较高的纯度。结论从人鼻粘膜取材,可以分离培养出OECs,体外生长增殖旺盛,并且经纯化后具有较高的纯度。

The Abstraction,cultivation and biological properties of OECs from the human olfactory mucosa

Objective To explore the Abstraction,cultivation,purification and verification of olfactory ensheathing cells(OECs) from the human olfactory mucosa,and observed their growth condition,so as to analyze their biological properties.We hoped to provides a viable method for patients with neurological disorders to implement the same kinds of cell transplantation.Methods Olfactory mucosa was obtained from 1/3 posterior part of patients’nasal septum who dead in traffic accident,unexpected diseases and patients who were made operation through sinus sphenoidalis with the permission of patients’dependents.Human OM-OECs were purified with the method of immuno-adsorption in culture dish coated with anti-p75,and examined by immunocytochemistry.Results Human OM-OECs prolifed at the most speed in vitro after 10 days.Both P75 and GFAP positive cells counted to above 90%.It demonstrated we could get human OM-OECs of hypso-purity.Conclusion OECs could obtain from patients’olfactory mucosa,prolife productively in vitro and hypso-purity could be achived by purification.