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目的建立稳定的人口腔颊粘膜上皮细胞体外培养方法。方法取正常人口腔颊粘膜,中性蛋白酶和胰蛋白酶消化后,获取的粘膜上皮细胞接种于10%含胎牛血清培养基中连续培养,探索最适培养条件,进行细胞定性及形态学观察。结果体外培养细胞中无成纤维细胞混杂,长满后呈上皮细胞特有的铺路石样外观,体外可连续传9~10代,生长期50~60天。细胞角蛋白免疫组化染色阳性,透射电镜下可见上皮细胞间特有的桥粒结构。结论采用含血清培养基体外连续培养口腔粘膜上皮细胞获得成功,4代以内培养上皮细胞形态结构与原代无差别,可用于组织工程化口腔粘膜构建。
Objective To establish a stable culture method of human buccal mucosal epithelial cells in vitro. Methods Normal human buccal mucosa, neutral protease and trypsin digestion, the obtained mucosal epithelial cells were inoculated in 10% fetal bovine serum-containing medium for continuous culture to explore the optimal culture conditions, qualitative and morphological observation of cells. Results In vitro cultured cells without fibroblasts mixed after overgrowth of epithelial cell-specific appearance of paving stones, in vitro continuous 9 to 10 generations, the growth period of 50 to 60 days. Cytokeratin immunohistochemical staining positive, transmission electron microscopy can be found in epithelial cells specific desmosomal structure. Conclusions Successive culture of oral mucosal epithelial cells with serum-containing medium is successful. The morphology and structure of cultured epithelial cells within 4 generations are indistinguishable from that of primary ones, and can be used to construct tissue-engineered oral mucosa.