酶消化组织块法培养人牙髓细胞

目的建立用酶消化组织块培养人牙髓细胞的方法。方法用酶消化组织块进行人牙髓细胞体外培养，倒置显微镜和透射电镜分别观察牙髓细胞形态、生长情况和超微结构，取第5代牙髓细胞测定生长曲线、免疫组化法鉴定组织来源，并检测其矿化能力。结果用酶消化组织块培养的人牙髓细胞成纤维细胞样，接种4天大部分组织块能有效贴附于培养瓶壁，第7天可见组织块边缘有细胞延展生长，第14天复层生长，呈网状，可顺利传代；牙髓细胞抗波形蛋白染色阳性，抗角蛋白染色阴性；细胞I型胶原染色阳性，连续培养的牙髓细胞可形成矿化结节，符合牙髓细胞的形态学特征和生物学特性。结论酶消化组织块培养人牙髓细胞方法简单可行，具有较高的成功率。

Culture of human dental pulp cells by explants with enzymatic digestion

Objective To establish the tissue black enzymolytic method in cultures of human dental pulp cells.Methods The human dental pulp cells were cultured with tissue-explants enzymatic digestion method.The morphological characters and uhrastructure of the cells were observed by inverted microscope and transmission electron microscope.The cells were immunohistochemically detected to identify the cell phenotype,the ability of mineralization.Results Cultures of human dental pulp cells were fibroblast-like and well-attached to wall 4 days after passage,and became growth after 7 days,muhilayered after 14 days.Anti-vimentin was positive,anti-cytokeratin was negative and type I collagen expression was positive by immunohistochemical staining.Cellular nodules were formed in human dental pulp cell after consecutive cuhure.Their morphological and biologic characteristics were similar to those of dental pulp cells.Conclusion The tissue black enzymolytic method in cultures of human dental pulp cells is simple and feasible.The method has a high success rate.