Expression Pattern of Oct-4, Sox2, and c-Myc in the Primary Culture of Human Dental Pulp Derived Cells
Expression Pattern of Oct-4, Sox2, and c-Myc in the Primary Culture of Human Dental Pulp Derived Cells
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Oct-4、Sox2 和 c-Myc 在人牙髓源细胞原代培养物中的表达模式
DOI：
10.1016/j.joen.2010.12.012
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发表时间：
2011-04-01
影响因子：
4.2
通讯作者：
Xiao, Yin
中科院分区：
文献类型：
--
作者：
Liu, Lu;Wei, Xi;Xiao, Yin
牙髓细胞在牙组织修复和再生方面具有广阔的应用前景。然而，在体外培养过程中，这些细胞经历了复制性衰老，导致细胞增殖和分化的显著改变。近年来，有报道称Oct-4、Sox2、c-Myc、Klf4等转录因子在干细胞自我更新即细胞重编程过程中发挥调控作用。因此，牙髓细胞在培养过程中的复制性衰老是否与这些转录因子的表达减少有关，这是一个有趣的问题。本研究通过免疫荧光染色和实时聚合酶链反应分析，研究了重编程标记物Oct-4、Sox2和c-Myc在体外外植体培养牙髓组织和外植体培养牙髓细胞（DPCs）中不同传代的表达情况。我们的结果表明，在外植体培养的DPCs中，Oct-4、Sox2和c-Myc在前2代传代后从细胞核转移到细胞质中。Oct-4、Sox2和c-Myc mRNA的表达量在前2代显著升高，在第2代达到峰值（P < 0.05），随后随着传代次数的增加而降低（P < 0.05）。我们首次证实了重编程标记Oct-4、Sox2和c-Myc在早期传代的DPC中被检测到，这些标记在DPC培养过程中在细胞核中的连续丢失可能与当前培养条件下牙髓源性细胞在长期体外培养过程中的细胞命运有关。（J end 2011;37:46 -472）
Dental pulp cells (DPCs) have shown promising potential in dental tissue repair and regeneration. However, during in vitro culture, these cells undergo replicative senescence and result in significant alteration in cell proliferation and differentiation. Recently, the transcription factors of Oct-4, Sox2, c-Myc, and Klf4 have been reported to play a regulatory role in the stem cell self-renewal process, namely cell reprogramming. Therefore, it is interesting to know whether the replicative senescence during the culture of dental pulp cells is related to the diminishing of the expression of these transcription factors. In this study, we investigated the expression of the reprogramming markers Oct-4, Sox2, and c-Myc in the in vitro explant cultured dental pulp tissues and explant cultured dental pulp cells (DPCs) at various passages by immunofluorescence staining and real-time polymerase chain reaction analysis. Our results demonstrated that Oct-4, Sox2, and c-Myc translocated from nucleus in the first 2 passages to cytoplasm after the third passage in explant cultured DPCs. The mRNA expression of Oct-4, Sox2, and c-Myc elevated significantly over the first 2 passages, peaked at second passage (P < .05), and then decreased along the number of passages afterwards (P < .05). For the first time we demonstrated that the expression of reprogramming markers Oct-4, Sox2, and c-Myc was detectable in the early passaged DPCs, and the sequential loss of these markers in the nucleus during DPC cultures might be related to the cell fate of dental pulp derived cells during the long-term in vitro cultivation under current culture conditions. (J Endod 2011;37:466-472)