论文部分内容阅读
目的探讨藓化饮对体外培养的人口腔黏膜上皮细胞增殖活性的影响。方法 DispaseⅡ及胰酶消化法原代培养人口腔黏膜上皮细胞,传代后HE染色,光镜下进行形态学观察。免疫组织化学抗角蛋白抗体染色,进行来源鉴定。取第二代培养细胞,随机分为4个实验组和1个对照组。制备藓化饮药液,实验组分别加入经无血清培基稀释为10~4、10~5、5×10~5、10~6μg/L4个浓度梯度的藓化饮液继续培养;对照组只加无血清培基继续培养。甲基噻唑基四唑法测定培养细胞的增殖活性。结果原代培养的人口腔黏膜上皮细胞呈扁平椭圆形或多角形,胞核清晰。细胞抗角蛋白抗体染色阳性。藓化饮在10~5μg/L浓度下增殖活性吸光度值明显高于其他浓度组和对照组(P<0.05)。而在10~4μg/L、5×10~5μg/L、10~6μg/L浓度下增殖活性吸光度值虽高于对照组,但差异无统计学意义(P>0.05)。结论使用无血清培养基可成功进行人口腔黏膜上皮细胞原代培养及传代培养。藓化饮在浓度为10~5μg/L下能明显增强体外培养的人口腔黏膜上皮细胞的增殖活性。
Objective To investigate the effect of moss beverage on human oral mucosal epithelial cell proliferation in vitro. Methods Human oral epithelial cells were primarily cultured by Dispase Ⅱ and trypsin digestion. HE staining was performed after passage and morphological changes were observed under light microscope. Immunohistochemistry Anti-keratin antibody staining for source identification. The second generation of cultured cells were randomly divided into 4 experimental groups and 1 control group. The mossy drink liquid was prepared and the experimental group was added with the mossy drink solution with 4 concentration gradients of 10 ~ 4, 10 ~ 5, 5 × 10 ~ 5 and 10 ~ 6μg / L diluted in the serum-free medium respectively. Only add serum-free medium to continue training. Methyl thiazolyl tetrazolium assay for determination of proliferative activity of cultured cells. Results Primary cultured human oral epithelial cells were flat oval or polygonal with clear nucleus. Cell anti-keratin antibody staining positive. The concentration of mossy drink at 10 ~ 5μg / L concentration of proliferative activity was significantly higher than other concentration and control groups (P <0.05). However, the proliferative activity absorbance values at 10-4μg / L, 5x10-5μg / L and 10-6μg / L were higher than those in the control group, but the difference was not statistically significant (P> 0.05). Conclusion The serum-free culture medium can be successfully used for primary culture and subculture of human oral epithelial cells. Mossy drink at a concentration of 10 ~ 5μg / L can significantly enhance the proliferation of cultured human oral mucosal epithelial cells.