摘要目的建立一个稳定的人正常口腔粘膜角化细胞体外培养体系.方法取正常口腔粘膜,经dispase及胰蛋白酶消化获取细胞,采用无血清培养液进行原代及传代培养,并进行形态学观察和角蛋白免疫组化染色等一系列细胞定性研究. 结果获取的细胞为单一上皮细胞;细胞可连续传4-5代,成活30-50天;细胞呈铺路石状,为上皮细胞的典型形态;电镜下见细胞具有丰富的张力丝和桥粒等特征性超微结构;角蛋白免疫组化染色阳性. 结论应用dispase酶及无血清培养液,在无3T3细胞的条件下可成功进行人正常口腔粘膜角化细胞连续传代培养.

abstractsAbstract:Objective To establish a method for culturing normal human oral keratinocytes.Methods Specimens obtained from healthy humans undergoing oral surgery were dissociated into single cell suspensions by dispase and trypsin. The cells were grown in serum-free medium. Morphological characteristics were studied under light microscope and electron microscope. Cytokeratins were shown by immunohistochemistry.Results Cells could be maintained in culture up to 4-5 passages or 30-50 days. Electron microscope revealed that there were desmosomes and tonofibrils in the oral keratinocytes. The cells showed positive staining for cytokeratin antibody. Conclusion Human oral keratinocytes have been successfully grown in serial culture.