Ameloblasts are specialized cells derived from the dental epithelium that produce enamel, a hierarchically structured tissue comprised of highly elongated hydroxylapatite (OHAp) crystallites. The unique function of the epithelial cells synthesizing crystallites and assembling them in a mechanically robust structure is not fully elucidated yet, partly due to limitations with in vitro experimental models. Herein, we demonstrate the ability to generate mineralizing dental epithelial organoids (DEOs) from adult dental epithelial stem cells (aDESCs) isolated from mouse incisor tissues. DEOs expressed ameloblast markers, could be maintained for more than five months (11 passages) in vitro in media containing modulators of Wnt, Egf, Bmp, Fgf and Notch signaling pathways, and were amenable to cryostorage. When transplanted underneath murine kidney capsules, organoids produced OHAp crystallites similar in composition, size, and shape to mineralized dental tissues, including some enamel-like elongated crystals. DEOs are thus a powerful in vitro model to study mineralization process by dental epithelium, which can pave the way to understanding amelogenesis and developing regenerative therapy of enamel.

成釉细胞是源自牙上皮的特殊细胞，可产生牙釉质，牙釉质是一种由高度细长的羟基磷灰石 (OHAp) 微晶组成的分层结构组织。上皮细胞合成微晶并将其组装成机械坚固结构的独特功能尚未完全阐明，部分原因是体外实验模型的限制。在此，我们证明了从小鼠门牙组织中分离出的成体牙上皮干细胞（aDESC）产生矿化牙上皮类器官（DEO）的能力。 DEO 表达成釉细胞标记，在含有 Wnt、Egf、Bmp、Fgf 和 Notch 信号通路调节剂的体外培养基中可以维持五个月以上（11 代），并且适合冷冻保存。当移植到小鼠肾囊下方时，类器官会产生 OHAp 微晶，其成分、大小和形状与矿化牙组织相似，包括一些牙釉质样细长晶体。因此，DEO 是研究牙上皮矿化过程的强大体外模型，可以为理解牙釉质生成和开发牙釉质再生疗法铺平道路。