人牙源性上皮细胞的体外培养研究

目的：建立人牙源性上皮细胞的体外培养方法，为牙齿组织工程研究提供可靠的种子细胞来源。方法：采用组织块法原代培养人牙源性上皮细胞，用更换培养液类型、多次差别消化法和反复贴壁法进行纯化，在倒置显微镜下观察细胞形态及生长状况，用免疫荧光法检测细胞表达角蛋白和成釉蛋白的情况。结果：原代培养的人牙源性上皮细胞生长良好，但有少量成纤维细胞混杂，经纯化后明显好转，上皮细胞呈片状生长，表达角蛋白及成釉蛋白。传至第5代后，细胞逐渐变为长梭形，失去上皮细胞形态。结论：体外培养的人牙源性上皮细胞在较长时间内可保持其特性，有望作为牙齿组织工程研究的种子细胞。

Study on in vitro culture of human dental epithelial cells

Objective:To develop a culture model for human dental epithelial cells and provide it as seed cells for tooth engineering. Method: Human dental epithelial cells were primarily obtained from tissue explant and purified using several methods combinedly. The cultured cells were examined for the expression of cytokeratin and ameloblastin by immunofluorescent staining.Result: The primary cultured human dental epithelial cells were mixed with a few of fibroblasts. But after purification, the dental epithelial cells had a morphology of typical epithelial cells and expressed cytokeratin and ameloblastin. The cells turned to be fibroblast-like cells when passaged to passage 5.Conclusion: Human dental epithelial cells could be cultured in vitro successfully and keep their biological characteristics for a relatively long time. It was hopeful that the cells would be used as seed cells for tooth engineering.