Differentiation of mouse-induced pluripotent stem cells into dental epithelial-like cells in the absence of added serum
Differentiation of mouse-induced pluripotent stem cells into dental epithelial-like cells in the absence of added serum
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DOI：
10.1007/s11626-019-00320-z
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发表时间：
2019-02-01
影响因子：
2.1
通讯作者：
Kato, Koichi
中科院分区：
文献类型：
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作者：
Abdullah, Aimi Naim;Miyauchi, Satoshi;Kato, Koichi
最近的研究通过模拟牙齿器官发生中牙上皮细胞和间充质细胞之间的相互作用，成功地生成了牙齿样结构。然而，这些方法的临床应用主要由于缺乏合适的牙上皮细胞来源而受到限制。诱导多能干细胞（iPSC）由于其独特的特性而作为牙上皮细胞的来源很有吸引力。在这项研究中，我们研究了神经营养蛋白-4 (NT-4) 对小鼠 iPSC (miPSC) 分化为牙上皮细胞的影响。我们的结果表明，在胚状体形成过程中添加NT-4显着触发了p63和CK14等上皮标记物的上调，表明NT-4为miPSC分化为上皮细胞提供了诱导条件。 NT-4处理的细胞在无血清培养条件下的扩增改善了具有鹅卵石样形态的细胞的形成，并显着下调了多能和外胚层标志物的表达。表型分析显示，牙上皮表面标记物 CD49f 在这些细胞上高度表达。通过成釉细胞特异性标记物的表达进一步证实了 miPSC 衍生的牙上皮样细胞的形成。这些结果表明，在拟胚体形成过程中添加NT-4以及无血清培养条件促进了miPSC向牙上皮样细胞的分化。
Recent studies have successfully generated tooth-like structure by mimicking the reciprocal interaction between dental epithelial and mesenchymal cells in tooth organogenesis. However, clinical applications of these methods are limited primarily due to the lack of appropriate sources for dental epithelial cells. Induced pluripotent stem cells (iPSCs) are attractive as a source for dental epithelial cells due to their unique characteristics. In this study, we examined the effect of neurotrophin-4 (NT-4) on the differentiation of mouse iPSCs (miPSCs) into dental epithelial cells. Our results showed that the addition of NT-4 during the formation of embryoid body significantly triggered the upregulation of epithelial markers such as p63 and CK14, suggesting that NT-4 provides an inductive condition for the differentiation of miPSCs into epithelial cells. Expansion of the NT-4-treated cells under serum-free culture conditions improves the formation of cells with cobblestone-like morphology and significantly downregulated the expression of pluripotent and ectodermal markers. Phenotypic analysis revealed that a dental epithelial surface marker, CD49f, was highly expressed on these cells. Formation of miPSCs-derived dental epithelial-like cells was further confirmed by the expression of ameloblast-specific markers. These results suggest that the addition of NT-4 during the formation of embryoid body together with the serum-free culture condition promoted the differentiation of miPSCs into dental epithelial-like cells.