兔口腔黏膜上皮细胞的体外培养方法

Primary culture in vitro of rabbit oral mucosal cells

目的培养兔口腔黏膜上皮细胞,建立稳定的口腔黏膜上皮细胞体外培养体系。方法取兔口腔黏膜组织,用0.25%中性蛋白酶(DispaseⅡ)分离上皮与皮下组织,采用无血清及无3T3细胞培养体系,即角质细胞培养液(DefinedK-SFM)培养兔口腔黏膜上皮细胞,并进行形态学观察及免疫细胞化学检测。结果用中性蛋白酶可成功分离上皮与皮下组织,原代培养11d细胞可融合成片,呈铺路石样,鉴定为单一口腔黏膜上皮细胞。结论利用角质细胞培养液可成功培养出兔口腔黏膜上皮细胞。

Objective To explore a new method for culturing rabbit oral mucosal epithelial cells in vitro. Methods Oral mucosal epithelial cells in rabbits were isolated by 0.25 % Dispase Ⅱ , and cultured by new culture system Defined K-SFM（ free serum medium and 3T3 cells）. The cultured cells were identified by immunohistochemistry. Results Rabbit oral mucosal epithelial tissue was obtained successfully by Dispase Ⅱ. After 11 d culture, the confluent cells were identified as pure oral muozsal epithelial cells. Conclusion The results indicate that the cells could be cultured successfully using Defined K-SFM.