Program in Craniofacial and Mesenchymal Biology and Department of Orofacial Sciences, University of California San Francisco, San Francisco, California, USA.
Tissue Eng Part C Methods. 2013 Jan;19(1):15-24. doi: 10.1089/ten.TEC.2012.0232. Epub 2012 Aug 16.
Dental epithelial stem cells (DESCs) drive continuous growth in the adult mouse incisors. To date, a robust system for the primary culture of these cells has not been reported, and little is known about the basic molecular architecture of these cells or the minimal extracellular scaffolding that is necessary to maintain the epithelial stem cell population in an undifferentiated state. We report a method of isolating DESCs from the cervical loop of the mouse mandibular incisor. Cells were viable in a two-dimensional culture system and did not demonstrate preferential proliferation when grown on top of various substrates. Characterization of these cells indicated that E-cadherin, integrin alpha-6, and integrin beta-4 mark the DESCs both in vivo and in vitro. We also grew these cells in a three-dimensional microenvironment and obtained spheres with an epithelial morphology and expression patterns. Insights into the mechanisms of stem cell maintenance in vitro will help lay the groundwork for the successful generation of bioengineered teeth from adult DESCs.
牙上皮干细胞（DESCs）驱动成年小鼠切牙的持续生长。迄今为止，尚未报道用于这些细胞的原代培养的稳健系统，并且对这些细胞的基本分子结构或维持上皮干细胞群体处于未分化状态所必需的最小细胞外支架知之甚少。我们报告了一种从下颌切牙颈环分离 DESCs 的方法。细胞在二维培养系统中具有活力，并且在各种基质上生长时不会表现出优先增殖。对这些细胞的表征表明，E-钙黏着蛋白、整合素 α6 和整合素 β4 在体内和体外均标记 DESCs。我们还在三维微环境中培养这些细胞，获得具有上皮形态和表达模式的球体。对体外干细胞维持机制的深入了解将有助于为从成年 DESCs 成功生成生物工程牙齿奠定基础。
