永生化人成牙本质细胞样细胞系的建立  

Establishment of immortalized human odontoblast-like cell line

作　　者：王捍国[1] 肖明振[1] 赵守亮[1] 郝建军[1] 张进[1] 

机构地区：[1]第四军医大学口腔医学院牙体病科,陕西西安710033

出　　处：《第四军医大学学报》2003年第10期876-878,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金 (39970 792 ;30 2 70 374) ;全军"十五"计划基金 (0 1 2 0 89) ;国家教育部回国人员启动基金 (2 0 0 0HG0 0 3)

摘　　要：目的 :建立永生化人成牙本质细胞系 .方法 :采用脂质体转染法 ,将人端粒酶逆转录酶 (hTERT)基因导入原代人成牙本质细胞样细胞 .培养液加G4 18筛选约 1mo后 ,抗性细胞克隆形成 ,滤纸片法转移、扩增并连续培养 .检测转染细胞内hTERT的基因整和和表达 ,初步分析细胞系的生物学特征 .结果 :hTERT稳定转染入目的细胞 ,表达mRNA及其蛋白 .转染后共获得 5个细胞克隆 ,克隆 5b来源的细胞在体外长期连续培养下生长良好 ,具有较高碱性磷酸酶活性 ,在转录水平表达成牙本质细胞特异性标志牙本质涎磷蛋白 ,细胞核型正常 .该细胞系至今传代 5 6代 ,约 6mo,命名为hTERT hOd l.结论AIM: To establish an immortalized human odontoblast cell line. METHODS: Primary human odontoblast like cells were cultured and transfected with human telomerase reverse transcriptase (hTERT) gene. Anti drug cell clones were formed by screening with G418, and were selected using filter paper and expanded for continuous culture. Integration and expression of hTERT in the transfected cells were detected and the biological characteristics of the cell line were analyzed. RESULTS: After stable transfection, mRNA and protein of hTERT were expressed in transfected cells. Among the 5 cell clones, cells derived from 5b showed high activity of alkaline phosphatase (ALP), normal phenotype and expression of dentin sialophosphoprotein (DSPP), which was the specific marker for odontoblast at mRNA level. The cell line had been in culture for about 6 months, 56 passages, which we named hTERT immortalized odontoblast like cell line (hTERT hOd l). CONCLUSION: An immortalized human odontoblast like cell line hTERT hOd l has been established.

关 键 词：成牙质细胞 永生化 细胞系 端粒 末端转移酶 

分 类 号：R780.2[医药卫生—口腔医学]