Development of technical bases for biotooth formation by using iPS cells

利用iPS细胞开发生物牙形成的技术基础

• 批准号: 21592500
• 负责人: FUJIWARA Naoki
• 金额: $ 2.91万
• 依托单位: Iwate Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21592500/
• 关键词: Biotooth; iPS細胞; 歯の再生

Induced pluripotent stem(iPS) cells are generated from somatic cells by the simultaneous introduction of several factors ; and they differentiate into the3embryonic germlayers with an extensive proliferative capacity. Recently, many researchers have reported that iPS cells can differentiate into different cell types, such as neurons, cardiac myocytes, and renal lineage cells, under appropriate conditions. Therefore, iPS cells have emerged as potential cell sources for tooth regeneration. In this study, we aimed to form biotooth by recombination of iPS cell-induced dental germ cells with mouse dental germ cells. First, we established the induction methods of iPS into mesenchymal stem cell(MSC). The differentiated cells expressed STRO-1, MSC marker, more than90%, as well as other MSC markers. Further, we established a culture protocol to induce the differentiation of iPS cells into neural crest like cells(NCLC), which is a precursor cell of dental mesenchymal cells. After the induction, the most of cells expressed neural crest cells markers. The cells did not form teratomas after transplantation into nude mice subcutaneously, suggesting the safety for clinical applications. Moreover, in recombination cultures of NCLC and mousedental epithelium, NCLC exhibited a gene expression pattern involving dental mesenchymal cells. Some NCLC also expressed dentin sialoprotein. Conditioned medium of mouse dental epithelium cultures further enhanced the differentiation of NCLC into odontoblasts. These results suggest that iPS cells are useful cell sources for tooth regeneration and tooth development studies.

诱导多能干细胞(IPS)是在多种因素的共同作用下，由体细胞分化为3个胚胎生殖层，具有广泛的增殖能力。最近，许多研究人员报道，iPS细胞在适当的条件下可以分化为不同类型的细胞，如神经元、心肌细胞和肾系细胞。因此，iPS细胞已成为牙齿再生的潜在细胞来源。在这项研究中，我们的目标是通过iPS细胞诱导的牙齿生殖细胞与小鼠牙齿生殖细胞的重组形成生物牙。首先，我们建立了诱导iPS分化为间充质干细胞的方法。分化后的细胞表达Stro-1、MSC标记物90%以上以及其他MSC标记物。此外，我们建立了诱导iPS细胞分化为神经脊样细胞(NCLC)的培养方案，NCLC是牙间充质细胞的前体细胞。诱导后，大部分细胞表达神经脊细胞标志物。细胞移植到裸鼠皮下后未形成畸胎瘤，提示临床应用是安全的。此外，在NCLC和鼠牙上皮的重组培养中，NCLC表现出涉及牙间充质细胞的基因表达模式。部分NCLC还表达牙本质唾液蛋白。小鼠牙上皮条件培养液进一步促进NCLC向成牙本质细胞分化。这些结果表明，iPS细胞是牙齿再生和牙齿发育研究的有用细胞来源。

项目成果

Live cell imaging of Hertwig's epithelial root sheath formation process

赫特维希上皮根鞘形成过程的活细胞成像

• 发表时间: 2010
• 作者: Hidemitsu Harada;Keishi Otsu
• 通讯作者: Keishi Otsu

Neural crest like cells from induced pluripotent stem cells for tooth regeneration

来自诱导多能干细胞的神经嵴样细胞用于牙齿再生

• 发表时间: 2010
• 作者: Ryota Kishigami;Keishi Otsu
• 通讯作者: Keishi Otsu

Hepatocyte growth factorはマウス臼歯歯胚の歯根形成を促進する

肝细胞生长因子促进小鼠磨牙牙根的形成

• 发表时间: 2011
• 作者: 藤原尚樹;櫻庭春菜;坂野深香;佐々木;及川愛;大津圭史;石関清人;原田英光
• 通讯作者: 原田英光

ライブイメージングでとらえる歯の発生メカニズム

通过实时成像捕捉牙齿发育机制

• 发表时间: 2012
• 作者: 大津圭史;原田英光
• 通讯作者: 原田英光

ヘルトヴィッヒ上皮鞘形成過程におけるサービカルループでの細胞動態

赫特维希上皮鞘形成过程中循环的细胞动力学

• 发表时间: 2009
• 作者: 原田英光;石関清人;大津圭史;鍵谷忠慶;藤原尚樹
• 通讯作者: 藤原尚樹