摘要
目的: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45颗,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(每法15颗)进行人牙髓细胞的体外原代培养,比较观察3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:3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,且呈成纤维细胞样,抗波形丝蛋白染色阳性,抗角蛋白染色阴性。其中,组织块法耗时较长,4例培养成功(26.7%);酶消化法培养成活率较低,且细胞数量较少,2例培养成功(13.3%);改良组织块法中未发现有漂浮的组织块,12例培养成功(80.0%)。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。
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 of human pulptissue cells.Immunocytochemical staining method was used to characterize the dental pulp cell lineage.Results： The cultured cells appeared to be fibroblast-like cells,which was positive for anti-vimentin and negative for anti-cytokeratin by immunohistochemical stain.The primary dental pulp cells cultured by the tissue explants method could reach confluence after 4 weeks and the culture success was 26.7%.The culture success of dental pulp cells was only 13.3% by collagenase/dispase digestion method.Dental pulp cells can be obtained by the modified tissue culture without floating tissue fragments and the success rate was 80.0%.Conclusion： Modified tissue culture techniques solved the difficulty of adherence of tissue fragments,simplified the procedure,and kept the high success rate.The modified tissue techniques culture established in this study is an effective method for the primary culture of human dental pulp cells in vitro.
出处
《天津医药》
CAS
北大核心
2011年第2期139-141,193,共4页
Tianjin Medical Journal
