Immortalization and characterization of human dental pulp cells with odontoblastic differentiation
Immortalization and characterization of human dental pulp cells with odontoblastic differentiation
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DOI：
10.1016/j.archoralbio.2007.02.006
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发表时间：
2007-08-01
影响因子：
3
通讯作者：
Takata, Takashi
中科院分区：
文献类型：
--
作者：
Kitagawa, Masae;Ueda, Hirota;Takata, Takashi
目的：体外培养生长稳定、矿化能力强的永生化人牙髓细胞。设计：从健康的第三磨牙中获得HDP细胞，通过转染人端粒酶转录酶（hTERT）基因使其永生化。为研究hTERT转染HDP细胞（HDP-hTERT）的特性，采用RT-PCR方法检测牙本质涎磷蛋白（DSSP）、Ⅰ型胶原（COLI）、碱性磷酸酶（ALP）和骨涎蛋白（BSP）mRNA的表达。此外，生化法检测ALP活性，茜素红S（ALZ）染色法检测细胞结节形成。hTERT基因。这些细胞绕过衰老，生长超过120个群体倍增（PDL），而没有显著的生长迟缓。RT-PCR证实hTERT在HDP-hTERT中高表达，并显示出明显的端粒酶活性。HDP-ori和HDP-hTERT均表达DSSP、COLI、ALP和BSP mRNA，ALP活性和ALZ染色均在同一水平。结论：本研究成功建立了人牙髓细胞向成牙本质细胞分化的永生化细胞系，为研究成牙本质细胞增殖和分化机制提供了一个有用的细胞模型。(C)2007爱思唯尔有限公司版权所有。
Objective: To immortalize human dental pulp (HDP) cell showing stable growth and high mineralization activities in vitro. Design: HDP cells were obtained from a healthy third molar and immortalized by transfection with human telomerase transcriptase (hTERT) gene. To examine the characters of hTERT transfected HDP (HDP-hTERT) cells, we examined expression of mRNA for dentin sialophosphoprotein (DSSP), type I collagen (COLI), alkaline phosphatase (ALP) and bone sialoprotein (BSP) by RT-PCR. In addition, we examined ALP activity by biochemical method and nodule formation by alizarin red S (ALZ) staining.Results: HDP-hTERT was obtained by transfection. with hTERT gene. These cells bypassed the senescence and grew over 120 population doublings (PDLs) without significant growth retardation. High expression of hTERT was confirmed in HDP-hTERT by RT-PCR and showed remarkable telomerase activity. Both HDP-original (HDP-ori) and HDP-hTERT expressed DSSP, COLI, ALP and BSP mRNA and showed ALP activity and ALZ staining at the same levels.Conclusions: We were able to establish a cell line of immortalized human dental pulp cells with odontoblastic differentiation which will be a useful cell model for studying the mechanism of proliferation and differentiation of odontoblasts. (C) 2007 Elsevier Ltd. All rights reserved.