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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 0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。
Objective To establish a stable human oral mucosal keratinocyte culture system in vitro. Methods Normal oral mucosa was obtained, and cells were obtained by dispase and trypsin digestion. Serum-free medium was used for primary and subculture. A series of cell qualitative studies including morphological observation and keratin immunohistochemical staining were performed. Results The cells obtained were single epithelial cells. The cells were passaged continuously for 4-5 passages and survived for 30-500 days. The cells were paved stone and were typical morphological epithelial cells. Under electron microscope, the cells were rich in tension filaments and desmosomes And other characteristic ultrastructure; keratin immunohistochemical staining. Conclusion Discontinuous subculture of human normal oral mucosa keratinocytes can be successfully performed in the absence of 3T3 cells using dispase enzyme and serum-free medium.