人牙槽成骨细胞的体外培养和生物学特性检测

In vitro culture and investigation of human osteoblasts from alveolar bone

目的：研究人牙槽成骨细胞体外培养及其生物学特性。方法：取人牙槽骨用组织块法进行体外细胞培养,用倒置显微镜观察细胞生长、细胞形态,噻唑蓝（MTT）比色法检测细胞增殖;在条件培养液中培养后用酶动力学方法作碱性磷酸酶（ALP）活性定量测定、Alizarin red染色和von Kossa染色作成骨特性检测,同位素3H掺入结合细菌胶原酶消化法测定I型胶原的合成。结果：培养第5天细胞从组织块中爬出,再培养14～16 d可以传代;在条件培养液中细胞ALP活性明显,Alizarin red染色和von Kossa染色细胞呈阳性,体外培养人牙槽成骨细胞具有分泌I型胶原的功能。结论：体外培养的人牙槽成骨细胞生长良好,并具有成骨细胞的形态和生物学特性。

Objective：To culture human alveolar osteoblasts and study its osteogenic characteristics in vitro.Methods：Human alveolar bone was obtained from human donor and cultured in explants.Cells migrating from explants were observed by inverted phase-contrast microscope and cell proliferation was detected by MTT assay.After cultured in conditional medium containing dexamethasone,β-glycerphosphoric sodium and ascorbic acid,cells were tested by ALP assay,Alizarin red assay and von Kossa assay,3H-proline labeling with collagenase digestion method.Results：The alveolar osteoblasts migrated from explants after 5 d culture.Cells could be passaged after 14-16 d culture.ALP values were obviously positive and cells showed positive reaction by Alizarin red assay and von Kossa assay in conditional medium.The cultured human alveolar osteoblasts secreted type I Collagen.Conclusion：The human alveolar osteoblasts cultured in this experiment grow well,and the morphological and biological characteristics of the culture cells are similar to those of osteoblasts.