Immortalization and characterization of human dental papilla cells with odontoblastic differentiation.
Immortalization and characterization of human dental papilla cells with odontoblastic differentiation.
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DOI：
10.1111/iej.12029
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发表时间：
2013-06
影响因子：
5
通讯作者：
Guobin Yang;X. Li;G. Yuan;Hailiang Liu;Mingwen Fan
中科院分区：
文献类型：
--
作者：
Guobin Yang;X. Li;G. Yuan;Hailiang Liu;Mingwen Fan
目的建立永生化人牙乳头细胞系。方法用含hTERT基因的慢病毒感染原代培养的hDPC。用聚合酶链式反应检测hTERT基因的整合和转录。对细胞的形态、增殖、矿化等特征进行了分析。用免疫组织化学和实时定量RT-PCR方法检测成牙本质细胞相关标志物ALP、DMP1、DLX3、OSX、DSP和Nestin的表达。结果hTERT基因整合入永生化细胞基因组DNA(hDPC-TERT)并转录成mRNA。经过长期培养，hDPC-TERT避开了衰老，增加了120多个种群倍增。HDPC-TERT细胞具有较高的增殖率，但保留了原代hDPC的表型特征，碱性磷酸酶活性和矿化活性也较高。此外，hDPC-TERT细胞在维持培养液中不表达DSP和Nestin，但在成牙本质细胞诱导后高表达DSP和Nestin。结论建立了一株具有成牙本质细胞分化潜能的永生化人牙乳头细胞系。该细胞系可作为研究成牙本质细胞分化启动机制的细胞模型。
AIM To establish a cell line of immortalized human dental papilla cells (hDPCs). METHODOLOGY Primary hDPCs were cultured and infected with lentivirus containing the hTERT gene. Integration and transcription of the hTERT gene were verified by PCR. The characteristics of the cells, such as morphology, proliferation and mineralization, were analysed. Also, the expression of odontoblastic-related markers including ALP, DMP1, DLX3, OSX, DSP and Nestin, was detected by immunohistochemistry and real-time RT-PCR. RESULTS hTERT gene was integrated into genomic DNA of immortalized cells (hDPC-TERT) and transcribed into mRNA. With long-time culture, hDPC-TERT bypassed senescence and grew over 120 population doublings. hDPC-TERT cells have a higher proliferation rate, but retain the phenotypic characteristics of the primary hDPCs, and so was ALP activity and mineralization activity. Furthermore, the hDPC-TERT cells express no DSP and Nestin with maintenance medium, but highly expressed DSP and Nestin after odontoblastic induction. CONCLUSIONS A line of immortalized human dental papilla cells, which remains in an undifferentiated state and has odontoblastic differentiation potential, was established. This cell line can be used as a cell model for studying the mechanism of the initiation of odontoblast differentiation.