Creating in vitro models of diseases of the pancreatic ductal compartment requires a comprehensive understanding of the developmental trajectories of pancreas-specific cell types. Here we report the single-cell characterization of the differentiation of pancreatic duct-like organoids (PDLOs) from human induced pluripotent stem cells (hiPSCs) on a microwell chip that facilitates the uniform aggregation and chemical induction of hiPSC-derived pancreatic progenitors. Using time-resolved single-cell transcriptional profiling and immunofluorescence imaging of the forming PDLOs, we identified differentiation routes from pancreatic progenitors through ductal intermediates to two types of mature duct-like cells and a few non-ductal cell types. PDLO subpopulations expressed either mucins or the cystic fibrosis transmembrane conductance regulator, and resembled human adult duct cells. We also used the chip to uncover ductal markers relevant to pancreatic carcinogenesis, and to establish PDLO co-cultures with stellate cells, which allowed for the study of epithelial–mesenchymal signalling. The PDLO microsystem could be used to establish patient-specific pancreatic duct models.

创建胰腺导管室疾病的体外模型需要全面了解胰腺特定细胞类型的发育轨迹。在这里，我们报告了胰管样类器官 (PDLO) 从人类诱导多能干细胞 (hiPSC) 在微孔芯片上分化的单细胞特征，该微孔芯片促进了 hiPSC 衍生的胰腺祖细胞的均匀聚集和化学诱导。使用形成的 PDLO 的时间分辨单细胞转录谱和免疫荧光成像，我们确定了从胰腺祖细胞通过导管中间体到两种类型的成熟导管样细胞和一些非导管细胞类型的分化途径。PDLO 亚群表达粘蛋白或囊性纤维化跨膜电导调节因子，并且类似于人类成人导管细胞。我们还使用该芯片来揭示与胰腺癌发生相关的导管标志物，并建立 PDLO 与星状细胞的共培养物，从而可以研究上皮-间充质信号传导。PDLO 微系统可用于建立患者特异性胰管模型。