AIM; To compare the successfulness and the growth of human caries dental pulp stem cells(hCDPSCs) cultured with three different methods. METHODS: Twenty-five normal and caries human third molars were collected, the dental pulp tissues were cultured by the tissue explant method, tissue-explan collagenase digestion method and the combination of explant method and enzymatic separation method, respectively. The adherence of the explants, the morphology and quantity of cells were observed under a phase-contrast microscope. Culture duration was recorded. The clones were identified by expression of Stro-1 and CD90 and the growth curve of normal DPSCs and CDPSCs was drawn. RESULTS; Human normal DPSCs and CDPSCs could be cultured by all the three methods. A large number of human normal DPSCs and CDPSCs were cultured by the com hined explant method and enzymatic separation method in a shorter time, and these cells exhibited more vitality and more different morphologies. The growth rate of CDPSCs was higher than that of normal DPSCs . CONCLISION: The improved combination of explant method and enzymatic separation method is an ideal method for the primary culture of hCDPSCs in vitro, it may provide a methodological foundation for studying the mechanism of the formation of the tertiary dentine when the tooth was damaged.%目的:比较3种方法培养人龋损牙髓干细胞的成功率和细胞生长状态,以探求人龋损牙髓干细胞的最佳培养方法.方法:取18～22岁成人新鲜正常和龋损离体第三磨牙各25个,采用组织块法、酶消化法、改良组织块酶消化法培养牙髓干细胞.通过倒置显微镜观察组织块的贴壁以及细胞的形态和数量,并记录培养所需时间；有限稀释法纯化牙髓干细胞,流式细胞仪检测正常和龋损牙髓干细胞表面标记物Stro-1、CD 90的表达情况,绘制正常和龋损牙髓干细胞生长曲线.结果:组织块法、酶消化法和改良组织块酶消化法均可以培养出人龋损牙髓干细胞,其中改良组织块酶消化法可以在短时间内获得数量多,状态好,形态多样的细胞.通过有限稀释法获得的龋损牙髓干细胞生长速率高于正常牙髓干细胞(P＜0.05).结论:改良组织块酶消化法是一种较为理想的原代培养人龋损牙髓干细胞的方法.
展开▼
