牙釉质是由釉柱细胞分泌的，是人体最坚硬的物质，其作为一种护盾保护着牙齿。然而，在超过90%的成年人中，牙釉质逐渐受损或部分丢失，并且由于萌发牙齿缺乏釉柱细胞，无法再生。在这里，我们使用单细胞组合索引RNA测序（sci-RNA-seq）建立了发育中人类牙齿的时空单细胞普查，识别了调控人类釉柱细胞分化过程的调控机制。我们鉴定了胎儿发育期间支持细胞与釉柱细胞之间的关键信号通路，并在人类诱导多能干细胞（iPSCs）诱导分化出的人类釉柱细胞中重现了这些发现。此外，我们还在三维（3D）器官样体系统中建立了釉发育不全的疾病模型，并展示了釉柱细胞向矿化结构的成熟过程。这些研究为未来的再生牙科学铺平了道路。版权所有 © 2023 Elsevier Inc. 发表。

Tooth enamel secreted by ameloblasts (AMs) is the hardest material in the human body, acting as a shield to protect the teeth. However, the enamel is gradually damaged or partially lost in over 90% of adults and cannot be regenerated due to a lack of ameloblasts in erupted teeth. Here, we use single-cell combinatorial indexing RNA sequencing (sci-RNA-seq) to establish a spatiotemporal single-cell census for the developing human tooth and identify regulatory mechanisms controlling the differentiation process of human ameloblasts. We identify key signaling pathways involved between the support cells and ameloblasts during fetal development and recapitulate those findings in human ameloblast in vitro differentiation from induced pluripotent stem cells (iPSCs). We furthermore develop a disease model of amelogenesis imperfecta in a three-dimensional (3D) organoid system and show AM maturation to mineralized structure in vivo. These studies pave the way for future regenerative dentistry.Copyright © 2023. Published by Elsevier Inc.