Helmholtz Pioneer Campus, Helmholtz Zentrum München, Neuherberg, Germany.
Department of Internal Medicine I, Ulm University Hospital, Ulm, Germany.
Nat Biomed Eng. 2021 Aug;5(8):897-913. doi: 10.1038/s41551-021-00757-2. Epub 2021 Jul 8.
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.
要构建胰腺导管区疾病的体外模型，需要全面了解胰腺特异性细胞类型的发育轨迹。在这里，我们报告了在微井芯片上从人诱导多能干细胞 (hiPSC) 分化胰腺类器官 (PDLO) 的单细胞特征，该芯片有助于 hiPSC 来源的胰腺祖细胞的均匀聚集和化学诱导。通过对形成的 PDLO 进行时分辨的单细胞转录组分析和免疫荧光成像，我们鉴定了从胰腺祖细胞通过导管中间细胞到两种成熟的导管样细胞和少数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剂，并且类似于成人胰腺导管细胞。我们还使用该芯片揭示了与胰腺癌发生相关的导管标记物，并建立了与星状细胞的 PDLO 共培养，这允许研究上皮-间充质信号。该 PDLO 微系统可用于建立患者特异性胰腺导管模型。
