改良的兔口腔黏膜上皮体外培养方法

Improved procedure for the culture of the rabbit oral mucosal epithelial cells in vitro

目的分析兔口腔黏膜上皮细胞(OMECs)体外培养影响因素,观察OMECs细胞生物学变化的特点并寻找促进体外增殖OMECs的方法。方法比较不同的表面消毒方法、不同的DispaseⅡ配制、不同的培养基、不同的钙离子浓度的K-SFM对OMECs体外增殖的影响。结果 OMECs用2%碘酊表面消毒比使用1%聚维碘酮消毒能明显减少微生物的污染;OMECs用K-SFM配制的DispaseⅡ消化后能够很好的贴壁,用PBS配制的DispaseⅡ消化后细胞不能贴壁;OMECs在两种浓度钙离子的K-SFM培养基中生长无统计学差别;OMECs在血清存在的情况下生长快,但容易分化为成纤维样细胞。结论使用2%碘酊消毒,K-SFM配制DispaseⅡ消化,K-SFM作为培养基,可体外获得较纯净的OMECs,细胞显示上皮细胞特征。

Objective To explore the biological characteristics of rabbit oral mucosal epithelial cells （OMECs） and the suitable procedure for the culture of OMECs. Methods OMECs were cultured by different surface disinfection, different Dispase Ⅱ preparation, different medium, different calciumion concentration of K-SFM medium. Results Microbial contamination of cell culture could significantly reduce with 2% iodine solution used for sterilization of oral cavity before cell culture. The OMECs digested with Dispase Ⅱ in K-SFM could attach to the culture flask quickly, the OMECs digested with Dispase Ⅱ in PBS rare attached to the culture flask. The proliferations of OMECs cultured in 0 or 0.09 mmol/L calcium concentrations of K-SFM medium showed no statistically significant. The OMECs grew fast with serum, but it was easy to differentiate into fibroblast-like cells. Conclusion OMECs could grow well and had the typical OMEC morphology by the procedure that described in this experiment.