【摘要】 目的建立成人鼻黏膜嗅鞘细胞的体外培养方法,探讨干细胞标志蛋白在人嗅鞘细胞的表达及其意义,为嗅鞘细胞自体移植治疗中枢神经系统损伤提供可行性研究资料。方法用鼻内镜自成人(鼻中隔偏曲伴黏膜肥厚者)鼻腔活体钳取部分嗅区黏膜,经胰酶消化制成细胞悬液。细胞接种于玻璃培养瓶,用含10%胎牛血清的F12培养基培养、传代、扩增。用免疫细胞化学或免疫荧光染色和免疫印迹方法检测SOX-2、SOX-10、Nestin、NCAM、NGFRp75和S100-β在第7代嗅鞘细胞的表达。结果成人嗅黏膜细胞经过多次传代培养后,嗅鞘细胞逐渐成为优势细胞,该细胞高表达干细胞标志蛋白和嗅鞘细胞标志蛋白。结论成人嗅黏膜细胞体外培养可获得高产量的许旺细胞(Schwann cells,Scs)样嗅鞘细胞,该细胞具有干细胞特性(stemness)和很强的增殖能力,可作为自体细胞移植治疗中枢神经系统损伤的种子细胞。更多还原

【Abstract】 OBJECTIVE To explore the stemness of the olfactory ensheathing cells(OECs) from nasal olfactory mucosa of adult human. METHODS The nasal olfactory mucous membrane was alive removed from adult human, the cells of the mucous membrane were culture in F12 medium contained 10% fetal bovine serum. The growth and morphologic characteristics of cultured cells were observed by staining of immunocytochemistry or immunofluorescent with antibodies against SOX-2, SOX-10, Nestin, NCAM, NGFRp75and S100-β. The expressions of the proteins in the OECs were detected by werstern blotting. RESULTS The ensheathing cells derived from olfactory mucous membrane could proliferate and maintain expressions of SOX-2,SOX-10,Nestin, NCAM, NGFRp75, S100-β in vitro. CONCLUSION The cultured cells from nasal olfactory mucosa of adult human contain Schwann cell-like olfactory ensheathing cells with stemness; After subculturing the cells several times, the olfactory ensheathing cells become pure gradually. The olfactory ensheathing cells may be used as an autotransplant cells to repair the injured central and peripheral nervous system.更多还原