Newly established cell lines from mouse oral epithelium regenerate teeth when combined with dental mesenchyme
Newly established cell lines from mouse oral epithelium regenerate teeth when combined with dental mesenchyme
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DOI：
10.1007/s11626-009-9265-7
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发表时间：
2010-05-01
影响因子：
2.1
通讯作者：
Tomooka, Yasuhiro
中科院分区：
文献类型：
--
作者：
Takahashi, Chiho;Yoshida, Hiroyuki;Tomooka, Yasuhiro
本研究旨在探讨口腔上皮细胞克隆系与牙胚间充质复合后能否分化为成釉细胞并再生牙齿。从胚胎第18天（E18）的p53缺陷型胎鼠的口腔上皮中建立了具有不同形态的克隆细胞系。该品系小鼠被证明是一个有用的来源，建立克隆和永生化的细胞系，从各种组织和在不同的发展阶段。在E18时，牙齿形态发生几乎完成，口腔上皮与牙齿上皮分离。在细胞系的RT-PCR分析中，检测到了粘膜上皮标志物（细胞角蛋白14），但在塑料培养皿上培养细胞时，未检测到成釉细胞标志物，例如釉原蛋白和成釉蛋白。它们在饲养层或胶原凝胶上培养1-3周后形成复层上皮，并在上层表达特异性分化标志物（CK 1-3），表明它们是口腔粘膜来源的。然后，用细胞系和胎磨牙间充质组织制备生物工程牙胚，并植入肾包膜下2-3周。6个细胞系中有5个细胞再生了天然牙的钙化结构。我们的研究结果表明，一些口腔上皮细胞在E18具有分化为成釉细胞的能力。此外，本研究建立的细胞系是研究牙齿器官发生和牙齿再生过程的有用模型。
The present study attempted to examine whether clonal cell lines of the oral epithelium can differentiate into ameloblasts and regenerate tooth when combined with dental germ mesenchyme. Clonal cell lines with a distinct morphology were established from the oral epithelium of p53-deficient fetal mice at embryonic day 18 (E18). The strain of mouse is shown to be a useful source for establishing clonal and immortalized cell lines from various tissues and at various stages of development. Tooth morphogenesis is almost completed and the oral epithelium is segregated from the dental epithelium at E18. In RT-PCR analysis of cell lines, mucosal epithelial markers (cytokeratin 14) were detected, but ameloblast markers such as amelogenin and ameloblastin were not detected when cells were cultured on plastic dish. They formed stratified epithelia and expressed a specific differentiation marker (CK13) in the upper layer when cultured on feeder layer or on collagen gel for 1-3 wk, demonstrating that they are of oral mucosa origin. Next, bioengineered tooth germs were prepared with cell lines and fetal molar mesenchymal tissues and implanted under kidney capsule for 2-3 wk. Five among six cell lines regenerated calcified structures as seen in natural tooth. Our results indicate that some oral epithelial cells at E18 possess the capability to differentiate into ameloblasts. Furthermore, cell lines established in the present study are useful models to study processes in tooth organogenesis and tooth regeneration.