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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。
OBJECTIVE: To establish a method for culturing human dental epithelial cells in vitro and to provide a reliable source of seed cells for dental tissue engineering. Methods: Human dental epithelial cells were primarily cultured by tissue block method. The culture medium was replaced by different digestion method and repeated adherent method. The morphology and growth of cells were observed under an inverted microscope. The immunofluorescence Detection of cells expressing keratin and ameloblasts. Results: Human primary cultured human dental epithelial cells grew well, but a small amount of fibroblasts were mixed. After purification, the proliferation of human dental epithelial cells was obviously improved. The epithelial cells grew in sheet form and expressed keratin and ameloblastin. After the 5th passage, the cells gradually became long fusiform and lost epithelial cell morphology. CONCLUSION: Human dental epithelial cells cultured in vitro can retain their characteristics over a long period of time and are expected to be used as seed cells in dental engineering.