体外连续培养人口腔粘膜角化细胞

Serial cultivation of normal human oral keratinocytes

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～ 5 0d ;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

Objective To establish a method of culturing normal human oral keratinocytes. Methods Specimens obtained from healthy humans undergoing oral surgery were dissociated by Dispase and trypsin into a single cell suspension. The cells were grown in serum free medium. Morphological characteristics were studied under light microscope and electron microscope (EM). Cytokeratins were showed by immunohistochemistry. Results The cells could be maintained in culture up to 4～5 passages, or 30～50 days. EM revealed that there were desmosomes and tonofibrils in the culture cells. The cells showed positive staining for cytokeratin antibody.Conclusions The culture technique for growing normal human oral keratinocytes has been established.