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Immortalized mouse dental papilla mesenchymal cells preserve odontoblastic phenotype and respond to bone morphogenetic protein 2

机译:永生化的小鼠牙乳头间充质细胞保留牙本质母细胞表型并响应骨形态发生蛋白2

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摘要

Odontogenesis is the result of the reciprocal interactions between epithelial–mesenchymal cells leading to terminally differentiated odontoblasts. This process from dental papilla mesenchymal cells to odontoblasts is regulated by a complex signaling pathway. When isolated from the developing tooth germs, odontoblasts quickly lose their potential to maintain the odontoblast-specific phenotype. Therefore, generation of an odontoblast-like cell line would be a good surrogate model for studying the dental mesenchymal cell differentiation into odontoblasts and the molecular events of dentin formation. In this study, immortalized dental papilla mesenchymal cell lines were generated from the first mouse mandibular molars at postnatal day 3 using pSV40. These transformed cells were characterized by RT-PCR, immunohistochemistry, Western blot, and analyzed for alkaline phosphatase activity and mineralization nodule formation. One of these immortalized cell lines, iMDP-3, displayed a high proliferation rate, but retained the genotypic and phenotypic characteristics similar to primary cells as determined by expression of tooth-specific markers and demonstrated the ability to differentiate and form mineralized nodules. Furthermore, iMDP-3 cells had high transfection efficiency as well as were inducible and responded to BMP2 stimulation. We conclude that the establishment of the stable murine dental papilla mesenchymal cell line might be used for studying the mechanisms of dental cell differentiation and dentin formation.
机译:牙本质发生是上皮间质细胞之间相互相互作用导致最终分化的成牙本质细胞的结果。从牙乳头间充质细胞到成牙本质细胞的过程受复杂的信号通路调节。当从发育中的牙胚中分离出来时,成牙本质细胞迅速丧失了维持成牙本质细胞特异性表型的潜力。因此,成牙本质细胞样细胞系的生成将是研究牙科间充质细胞分化为成牙本质细胞和牙本质形成的分子事件的良好替代模型。在这项研究中,使用pSV40从出生后第3天的第一只小鼠下颌磨牙产生了永生化的牙乳头间充质细胞系。通过RT-PCR,免疫组织化学,蛋白质印迹对这些转化的细胞进行表征,并分析其碱性磷酸酶活性和矿化结节的形成。这些永生的细胞系之一iMDP-3表现出高增殖率,但保留了与原代细胞相似的基因型和表型特征,这是通过牙齿特异性标记物的表达确定的,并显示出分化和形成矿化结节的能力。此外,iMDP-3细胞具有很高的转染效率,并且可诱导并响应BMP2刺激。我们得出结论,建立稳定的鼠科牙乳头间充质细胞系可能用于研究牙细胞分化和牙本质形成的机制。

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