GSE162547
Single-cell-resolved differentiation of human induced pluripotent stem cells into pancreatic duct-like organoids on a microwell chip
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 facili-tates 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 pan-creas  progenitors  through  ductal  intermediates  to  two  types  of  mature  duct-like  cell  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. Overall design: single cell RNA sequencing of pancreatic duct-like organoids differentation kinetics