Characterization of human pancreatic tissue and organoids reveals subpopulation of intercalated duct cells with progenitor traits

Pancreatic duct organoids have emerged as a valuable tool for investigating pancreas biology and physiology through in vitro modeling. Despite their widespread use, a comprehensive understanding of the cellular heterogeneity of organoids and their underlying tissue remains elusive. Here we performed single-cell transcriptomic profiling of human adult pancreatic exocrine tissue and their derived organoids, revealing a gradient of keratin expression within the ductal network that clustered cells into two distinct low- and high-keratin subpopulations. Keratin-high cells display a higher capacity to form organoids while keratin-low cells express stemness associated markers. Finally, comparative gene expression analysis between ductal tissue and derived organoids revealed a partial recapitulation of the in vivo heterogeneity. These findings further our knowledge on pancreatic duct organoids and pave the way for future investigations into the heterogeneity of the pancreatic ductal syste Pancreatic islet-depleted tissue of 5 donors and 2 organoid lines was sequenced utilizing the SORT-seq platform. Samples of 2 donors were additionally collected using Fluorescence activated cell sorting based on expression of cell surface marker GP2 to study both positive and negative populations.