【摘要】 目的 :研究不同培养条件下口腔黏膜上皮细胞的生长情况 ,了解不同添加成分对上皮细胞生长和分化的影响 ,从而探索一条简便有效的培养口腔黏膜上皮细胞的方法。方法 :选取健康成年人拔除第三磨牙时切除的牙龈黏膜 ,在超净台内用眼科剪、镊尽可能去除上皮下结缔组织 ,滴加Dispase ,4℃冷消化 1 6～ 1 8h ,取上皮皮片用眼科剪剪成约 1mm3 大小 ,随机分成 1 2组 ,分别接种于 2 5mm培养瓶中 ,每瓶加入培养液 1ml(MCDB1 5 3) ,放入 37℃ ,体积分数为 5 %CO2 细胞培养箱中培养。 1 2组培养液中分别加入不同浓度的胎牛血清、表皮生长因子、胰岛素、氢化可的松、霍乱毒素和转铁蛋白 ,观察细胞生长情况。选取能够正常增殖的细胞进行传代培养 ,并于传代后的第 2d、4d、6d、8d、1 0d、1 2d、1 4d纪录细胞总数。结果 :低血清加入各种生长因子的培养液中上皮细胞生长良好 ,光镜下观察为典型的“铺路石”状 ,传代后经免疫组化证实是上皮细胞 ,而高血清培养液中由于成纤维细胞的生长导致无法传代。结论 :MCDB1 5 3培养液中加入胰岛素、氢化可的松、表皮生长因子和霍乱毒素所构成的培养液 ,是一种有效的口腔黏膜上皮细胞培养液更多还原

【Abstract】 Aim: To study the human oral mucosa epithelial cells growth requirements and the expression of differentiated characteristics and the effects of some growth factors and to seek a simple and optional medium for culturing the normal human oral mucosa epithelial cells.Methods: Specimens were obtained from healthy gingival overlying impacted third molar teeth of adult human. Dispase were used to separate epithelium from the underlying fibrous connective tissue. The epithelium were seeded into 12 dishes in 1ml MCDB153 medium.Cells were maintained in a humidified 37 ℃ incubator with 5% CO 2. Epidermal growth factor, serum, hydrocortisone, insulin, transferrin, and cholera toxin were added into different medium. The number of cells per dish was counted at day 2,4,6,8,10,12,14.Results: The epithelial cells in 0.5% serum had a higher proliferation speed and perfect differentiation while the cells in high serum were terminated because of overgrowthing fibroblasts.Conclusion: The MCDB153 with 0.5%serum, EGF, insulin, hydrocortisone, and cholera toxin is a simple and viable medium for culturing human oral mucosa epithelial cells.更多还原