Differentiation of Odontoblast-Like Cells From Mouse Induced Pluripotent Stem Cells by Pax9 and Bmp4 Transfection
Differentiation of Odontoblast-Like Cells From Mouse Induced Pluripotent Stem Cells by Pax9 and Bmp4 Transfection
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DOI：
10.5966/sctm.2014-0292
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发表时间：
2015-09-01
影响因子：
6
通讯作者：
Takano-Yamamoto, Teruko
中科院分区：
文献类型：
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作者：
Seki, Daisuke;Takeshita, Nobuo;Takano-Yamamoto, Teruko
近年来，牙齿再生领域取得了进展，人类牙齿再生在未来可能变得可行。由于诱导多能干细胞（iPS）在适当的条件下可以分化为牙源性细胞，iPS细胞是牙齿再生的潜在细胞来源。然而，诱导iPS细胞衍生的牙源性细胞的确定方法尚未建立。我们描述了一种利用基因转染从iPS细胞分化成牙本质细胞的新方法。我们产生了小鼠iPS细胞衍生的神经嵴样细胞（iNCLC），其表现出神经嵴标志物。接下来，我们通过转染Pax9和Bmp4表达质粒将iNCLC分化为成牙本质细胞样细胞。外源性Pax9上调iNCLC中Msx1和牙本质基质蛋白1（Dmp 1）的表达，但不上调骨形态发生蛋白4（Bmp 4）和牙本质唾液磷蛋白（Dspp）的表达。外源性Bmp4上调了iNCLC中Msx1、Dmp1和Dspp的表达，但不上调Pax9。此外，Pax9和Bmp4质粒共转染iNCLC中显示Pax9的表达高于单独使用Pax9质粒时。相反，外源性Pax9下调Bmp4过表达。Pax9和Bmp4共转染协同上调Dmp1表达，然而，Pax9过表达下调外源性Bmp4诱导的Dspp表达。总之，这些研究结果表明，外源性Pax9和Bmp4诱导的信号之间的相互作用调制Dmp1和Dspp的表达。总之，Pax9和Bmp4表达质粒转染iNCLC诱导与成牙本质细胞分化相关的基因表达，表明iNCLC分化为成牙本质细胞样细胞。iPS细胞来源的成牙本质细胞样细胞可能是牙齿再生的有用细胞来源。
The field of tooth regeneration has progressed in recent years, and human tooth regeneration could become viable in the future. Because induced pluripotent stem (iPS) cells can differentiate into odontogenic cells given appropriate conditions, iPS cells are a potential cell source for tooth regeneration. However, a definitive method to induce iPS cell-derived odontogenic cells has not been established. We describe a novel method of odontoblast differentiation from iPS cells using gene transfection. We generated mouse iPS cell-derived neural crest-like cells (iNCLCs), which exhibited neural crest markers. Next, we differentiated iNCLCs into odontoblast-like cells by transfection of Pax9 and Bmp4 expression plasmids. Exogenous Pax9 upregulated expression of Msx1 and dentin matrix protein 1(Dmp1) in iNCLCs but not bone morphogenetic protein 4 (Bmp4) or dentin sialophosphoprotein (Dspp). Exogenous Bmp4 upregulated expression of Msx1, Dmp1, and Dspp in iNCLCs, but not Pax9. Moreover, cotransfection of Pax9 and Bmp4 plasmids in iNCLCs revealed a higher expression of Pax9 than when Pax9 plasmid was used alone. In contrast, exogenous Pax9 downregulated Bmp4 overexpression. Cotransfection of Pax9 and Bmp4 synergistically upregulated Dmp1 expression; however, Pax9 overexpression downregulated exogenous Bmp4-induced Dspp expression. Together, these findings suggest that an interaction between exogenous Pax9- and Bmp4-induced signaling modulated Dmp1 and Dspp expression. In conclusion, transfection of Pax9 and Bmp4 expression plasmids in iNCLCs induced gene expression associated with odontoblast differentiation, suggesting that iNCLCs differentiated into odontoblast-like cells. The iPS cell-derived odontoblast-like cells could be a useful cell source for tooth regeneration.