不同原代培养方法培养人牙髓细胞的研究

目的：比较并建立一种操作简单方便的人牙髓细胞原代培养方法。方法：选择因正畸拔除的完整无龋双尖牙，无菌条件下取出牙髓，分别采用组织块法、酶消化法和改良组织块法进行人牙髓细胞的体外原代培养，比较观察三种不同培养方法的成活率、细胞形态和数量，测定生长曲线并进行细胞来源鉴定。结果：三种方法均可从人牙髓组织分离培养获得外胚间充质来源的牙髓细胞，呈成纤维细胞样，抗波形丝蛋白染色阳性，抗角蛋白染色阴性。组织块法耗时较长，培养成功率26.7%；酶消化法培养成活率较低,且细胞数量较少,培养成功率仅为13.3%；在改良组织块法中未发现有漂浮的组织块，培养成活率较高为80%。结论：改良组织块法解决了传统组织块法中组织块难以贴壁的问题，简化了操作步骤并具有较高的成功率,是一种科学可行的人牙髓细胞原代培养方法。

Study on Different Methods in Primary Cultures of Human Dental Pulp Cells in Vitro

Objective: To establish a simple primary culture technique of human dental pulp cells in vitro.Methods: The pulp tissue was separated from 45 healthy young human teeth extracted for orthodontic purpose.The pulp explants were cultured by three methods,tissue explants method,tissue-explants collagenase/dispase digestion method and modified tissue culture techniques respectively.The cell culture efficiency,cell morphology and quantity were observed and evaluated by phase-contract microscopy and growth curres o...