以L929细胞为饲细胞的口腔粘膜上皮细胞体外培养

A Study of Oral Epithelial Cells Cultured with L929 Cells as Seed Cells in vitro

目的 :探索口腔粘膜上皮细胞体外培养的技术和方法 ,为进一步采用组织工程技术构建口腔粘膜组织奠定基础。方法 :取刚离乳的新西兰幼兔口腔粘膜组织小块 ,酶消化成单细胞悬液 ,接种后以L92 9细胞为饲细胞培养 ,定期换液、传代。以相差显微镜每天动态观察细胞形态变化及生长增殖情况 ,细胞进行常规HE染色、免疫组化染色及流式细胞仪检查 ,并以扫描电镜、透射电镜观察其超微结构。结果 :整个上皮细胞生长期内无成纤维细胞混杂生长 ,均为单一的上皮细胞 ,并证实为二倍体细胞。细胞可传 11～ 13代 ,成活 5 0～ 6 0d。结论 :新西兰幼兔口腔粘膜上皮细胞可在体外进行培养 ,在一定时间内保持增殖能力。这不仅为组织工程化口腔粘膜构建奠定了基础 。

Objective:The purpose of study aimed at investigating the technique of culturing oral epithelia in vitro and to set up an experimental model for further reconstructing oral mucosa in vitro.Methods: The oral mucosa was taken from young New Zealand rabbits, and the mucosa was digested with enzyme and suspended in liquid to form cellular suspension. Being seeded, the cells were cultured motionlessly. The medium was changed regularly and the cells were subcultured.Results: The cultured cells were all epithelial cells without fibroblasts, and they were proved to be diploid cells. The cells were subcultured in 1～13 generations which survived for 50～60 days. Conclusion: The oral epithelium of young New Zealand rabbit can be cultured in vitro, maintaining the ability to proliferate in a certain period. It is a pilot study to reconstruct oral tissue in vitro.