人牙髓细胞的体外培养及生物学特性研究

The characteristic of human dental pulp cells cultured in vitro

目的:从年轻恒牙的牙髓组织中分离培养牙髓细胞,研究其表型及生物学特性。方法:选取因正畸或阻生拔除的年轻健康双尖牙或第三磨牙,取出牙髓,组织块酶消化法分离培养人牙髓细胞,免疫细胞化学检测第3代人牙髓细胞表面标志,并对体外培养的人牙髓细胞的矿化能力进行研究。结果:体外培养的人牙髓细胞表达Ⅰ型胶原及波形丝蛋白,少数细胞增殖可形成克隆并表达间充质干细胞的表面标志STRO-1。体外连续培养可形成钙化结节,加入矿化诱导液后钙化结节形成时间明显提前,牙髓细胞碱性磷酸酶(Alkaline phosphatase,ALP)活性增高。结论:人牙髓细胞中含有少量干细胞,体外培养的人牙髓细胞可向成牙本质细胞分化并形成钙化结节,体外矿化诱导可促进牙髓细胞向成牙本质细胞分化。

Objective：To study the characteristic of human dental pulp cells from permanent teeth of young people.Methods：The healthy teeth were selected from the young people because of orthodontics or obstruction.The dental pulp cells were separated and cultured by tissue block enzyme digestion method.Immunohistochemical studies were performed to characterize the pulp cells,and the capacity of mineralization was researched.Results： Collagen type I and Vimentin were expressed in human pulp cells.Colony and STRO-1 were expressed only in several cells.With continued culture,human dental pulp cells formed cell nodus.Applied with odontogenic induction medium,the time of cell nodus was advanced,and alkaline phosphatase activity was increased（P0.05）.Conclusion： There are only a few of human dental pulp stem cells in dental pulp.The continued culture of pulp cells has odontogenic differentiation.