人口腔黏膜上皮细胞的培养

目的:培养人口腔黏膜上皮细胞,建立稳定的人正常口腔黏膜角化细胞体外培养体系,为进一步采用组织工程技术构建口腔黏膜组织奠定基础。方法:取3个月唇裂患儿手术多余口腔黏膜,经Dispase及胰蛋白酶消化获取细胞,采用无血清角化细胞培养液进行原代培养,并进行形态学观察和角蛋白免疫组化染色等细胞定性研究。结果:在0.25×10~2个细胞的低密度接种下,经过20天培养,获得了完全融合成片的口腔黏膜原代细胞,细胞呈铺路石样生长,经鉴定为单一的口腔黏膜上皮细胞。结论:正常口腔黏膜组织经Dispase及胰蛋白酶消化,应用无血清培养液进行培养,可成功培养出人正常口腔黏膜上皮原代细胞。

Original Generation Cultivation of Normal Human Oral Keratinocytes

Objective: To cultivate normal human oral keratinocytes and lay a foundation for oral tissue-engineered mucosa. Methods: Specimens obtained from a healthy child undergoing oral surgery were dissociated by dispase and trypsin into a single cell suspension. The cells were grown in serum-free medium. Morphological characteristics were studied under light microscope and electron microscope. Results: After 20 days' culture, the monolayer confluent cells of normal human original generation oral keratinocytes were obtained. Conclusion: The culture technique for growing normal human oral keratinocytes has been established.