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.

通过同时引入几个因素，从体细胞中产生了诱导的多能干（IPS）细胞。它们以广泛的增殖能力分化为3embryonic系列。最近，许多研究人员报告说，在适当条件下，IPS细胞可以分化为不同的细胞类型，例如神经元，心肌细胞和肾脏谱系细胞。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，我们旨在通过将IPS细胞诱导的牙科细胞与小鼠牙科细胞重组形成生物齿。首先，我们将IPS的诱导方法建立到间充质干细胞（MSC）中。分化的细胞表达stro-1，MSC标记，超过90％以及其他MSC标记。此外，我们建立了一种培养方案，以诱导IPS细胞分化为神经rest（如细胞）（NCLC），该细胞（NCLC）是牙间充质细胞的前体细胞。诱导后，大多数细胞表达了神经rest细胞标记。细胞在皮下移植到裸鼠后没有形成畸胎瘤，这表明临床应用的安全性。此外，在NCLC和池化上皮的重组培养物中，NCLC表现出涉及牙科间充质细胞的基因表达模式。一些NCLC也表示牙本质唾液蛋白。小鼠牙科上皮培养的条件培养基进一步增强了NCLC为odontoblasts的分化。这些结果表明，IPS细胞是用于牙齿再生和牙齿发育研究的有用细胞来源。

项目成果

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

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

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