牙髓培养细胞体外诱导矿化能力：冠髓及根髓比较.doc

牙髓培养细胞体外诱导矿化能力：冠髓及根髓比较 　【摘要】 目的 比较人牙髓的根、冠髓培养细胞体外诱导矿化能力，探讨牙髓干细胞在牙髓中的定位。方法 用组织块法对人牙的根髓和冠髓细胞进行原代和传代培养，将第五代培养细胞用条件培养液（20％DMEM＋10nmol/L地塞米松＋10mmol/L β-甘油酸钠＋50mg/L L-抗坏血酸）诱导矿化，在诱导后第10、20、30天对细胞进行Von Kossa钙染色。结果 根髓中形成的矿化小结比冠髓中更早、更强烈。结论 根髓比冠髓更容易诱导矿化。牙髓干细胞可能存在于根髓和冠髓之中，在根髓中比冠髓中具有更高的密度。 【关键词】 牙髓干细胞；细胞培养；矿化结节 The induced mineralization ability in cultured dental pulp cells:the comparation between coronal and root pulp cells 【Abstract】 Objective To compare the mineralization ability between cultured dental root and coronal pulp cells,and to investigate the localization of dental pulp stem cells(DPSCs) in dental pulps. Methods The human dental root and coronal pulp cells were cultured in tissue-explant method,the 5thgeneration of cells were subcultured with conditioning medium(20%DMEM+10nmol/L +dexamethasone+50mg/L ascorbic acid) to induce mineralization. At the days of 10,20,30 after inducing,von Kossa for calcium staining was used to investigate the mineralization ability in root and coronal pulp cells. Results The mineralization nodules appeared earlier and stronger in the root pulp cells than that in the coronal pulp cells observed in phase-contrast microscope and von Kossa staining. Conclusion Root dental pulp cells could be induced to mineralization more easily than the coronal pulp cells . DPSCs may exist in both root and coronal pulp, and its density in the root pulp is higher than that in coronal pulp. 【Key words】 dental pulp stem cells；cell culture；mineralization nodules 牙髓干细胞（dental pulp stem cells，DPSCs）在成体牙髓腔中的定位问题是牙体组织工程学中尚未解决的基本问题。研究表明，体外培养的DPSCs和骨髓干细胞一样，都具有在一定条件下诱导矿化形成矿化小结的能力，矿化小结的形成是DPSCs分化的标记。本文通过对体外培养的人牙髓髓细胞诱导矿化，并对其矿化能力进行比较，以探讨DPSCs在牙髓中的定位。 1 材料与方法 1.1 主要实验器材及试剂 CO2培养箱（Nuaire, America）；超净工作台（苏净集团苏州安泰空气技术有限公司）；相差显微镜（Olympus, Japan ）；细胞培养瓶及培养板（Costar, America）；高速离心机（上海医用设备厂）；细胞记数板；高糖DMEM培养基（Gibco, USA）；胎牛血清（Hyclone公司）；地塞米松（Sigma, USA）；L-抗坏血酸（Amesco, USA）；β-甘油磷酸钠（E.Merck, Darmstadt）。 1.2 实验方法 1.2.1 细胞培养 收集5颗16～25岁健康人因正畸或阻生而拔出的第三磨牙，在无菌条件下分别取其根髓和冠髓，用组织块法进行细胞