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作 者:王静[1] 吴军正[1] 郭富平[1] 董纪军[1] 魏红宇[1]
机构地区:[1]西安第四军医大学口腔医学院口腔生物教研室,710032
出 处:《实用口腔医学杂志》2005年第5期587-591,共5页Journal of Practical Stomatology
基 金:国家自然科学基金;项目批准号:30371551
摘 要:目的建立体外培养人涎腺上皮细胞的方法,观察其体外生长的生物学特性。方法应用组织块培养法,用无血清培养基(SFM),无血清1∶1DMEM/F12以及含25ml/L胎牛血清的1∶1DMEM/F123种不同的培养基对10例人涎腺标本进行体外原代和传代培养。用倒置显微镜观察培养细胞体外生长的形态特征。用细胞计数法观察细胞生长情况。用HE染色、PAS染色、细胞角蛋白、Vimentin免疫组化染色对培养细胞进行形态学检查和鉴定。结果10例人涎腺标本在体外原代培养均有细胞长出,培养的细胞为上皮样。用SFM培养的细胞增殖较旺盛,可传代培养5次,用其它2种培养基培养的细胞仅可传代1次。细胞角蛋白染色阳性,Vi-mentin染色阴性,PAS染色阳性。结论应用组织块培养的方法可以获得原代和传代培养的人涎腺上皮细胞,SFM较1∶1DMEM/F12更适宜涎腺上皮细胞生长。Objective :To establish a culture method of human salivary gland epithelial cells and to study their growth characteristics in vitro. Methods:Tissue explant technic was employed to culture human salivary gland epithelial cells in serum free medium (SFM) ,1:1 DMEM/FI2 and 1:1 DMEM/F12 containing 25 ml/L fetal boven serum (FBS) respectively. The morphology of the cultured cells was observed by phase contract microscope. The cell growth was studied by cell counting. The cells were identified by HE staining, PAS staining and SP staining. Results: Growth of human salivary gland epithelial cells was observed in primary culture in the three kinds of medium in all 10 cases. The cultured cells were epidermoid, positive for cytokeratin, negative for Vimentin and positive for PAS staining. The cells in SFM could be subcultured for five passages,while only for one passage in the other two kinds of medium. Conclusion :SFM is superior to serum free 1:1 DMEM/F12 or 1:1 DMEM/F12 containing 25 ml/L FBS for the culture of hu- man salivary gland epithelial cells.
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