Induction of dental epithelial cell differentiation marker gene expression in non-odontogenic human keratinocytes by transfection with thymosin beta 4
Induction of dental epithelial cell differentiation marker gene expression in non-odontogenic human keratinocytes by transfection with thymosin beta 4
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DOI：
10.1016/j.scr.2013.11.006
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发表时间：
2014-01-01
影响因子：
1.2
通讯作者：
Sakai, Hidetaka
中科院分区：
文献类型：
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作者：
Kiyoshima, Tamotsu;Fujiwara, Hiroaki;Sakai, Hidetaka
先前的研究表明，从发育中的牙胚中释放出来的细胞的重组会发育成牙齿。然而，由于伦理问题，很难将人类发育中的牙胚用作细胞来源。研究表明，胸腺素β 4（Tmsb4x）与牙胚的发生和发育密切相关。我们在此尝试通过转染TMSB 4X从非牙源性HaCaT细胞建立牙源性上皮细胞。TMSB4X转染的细胞在钙化诱导培养基中培养时形成了茜素红S（ALZ）和von Kossa染色（磷酸钙沉积物）阳性的结节。显示出比其它克隆更大量的钙沉积的三个选择的克隆表达PITX 2、细胞角蛋白14和Sonic Hedgehog。牙胚发育相关基因RUNX2、Amelogenin、Ameloblastin和Enamelin表达上调。这些蛋白质在ALZ和von Kossa染色阳性的结节中进行免疫组化观察。RUNX2阳性选择的TMSB 4X转染细胞植入裸鼠背部皮下组织形成基质沉积。免疫组化染色显示，AMELX、AMBN和ENAM在基质沉积物中可见。本研究证明了TMSB 4X诱导非牙源性HaCaT细胞中牙上皮细胞分化标记基因表达的可能性。(C)2013作者Elsevier B.V.出版，保留所有权利。
Previous studies have shown that the recombination of cells liberated from developing tooth germs develop into teeth. However, it is difficult to use human developing tooth germ as a source of cells because of ethical issues. Previous studies have reported that thymosin beta 4 (Tmsb4x) is closely related to the initiation and development of the tooth germ. We herein attempted to establish odontogenic epithelial cells from non-odontogenic HaCaT cells by transfection with TMSB4X. TMSB4X-transfected cells formed nodules that were positive for Alizarin-red S (ALZ) and von Kossa staining (calcium phosphate deposits) when cultured in calcification-inducing medium. Three selected clones showing larger amounts of calcium deposits than the other clones, expressed PITX2, Cytokeratin 14, and Sonic Hedgehog. The upregulation of odontogenesis-related genes, such as runt-related transcription factor 2 (RUNX2), Amelogenin (AMELX), Ameloblastin (AMBN) and Enamelin (ENAM) was also detected. These proteins were immunohistochemically observed in nodules positive for the ALZ and von Kossa staining. RUNX2-positive selected TMSB4X-transfected cells implanted into the dorsal subcutaneous tissue of nude mice formed matrix deposits. Immunohistochemically, AMELX, AMBN and ENAM were observed in the matrix deposits. This study demonstrated the possibility of induction of dental epithelial cell differentiation marker gene expression in non-odontogenic HaCaT cells by TMSB4X. (C) 2013 The Authors. Published by Elsevier B.V. All rights reserved.