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  • Single-cell-resolved differentiation of human induced pluripotent stem cells into pancreatic duct-like organoids on a microwell chip.

Single-cell-resolved differentiation of human induced pluripotent stem cells into pancreatic duct-like organoids on a microwell chip.

Nature biomedical engineering (2021-07-10)
Sandra Wiedenmann, Markus Breunig, Jessica Merkle, Christine von Toerne, Tihomir Georgiev, Michel Moussus, Lucas Schulte, Thomas Seufferlein, Michael Sterr, Heiko Lickert, Stephanie Ellen Weissinger, Peter Möller, Stefanie M Hauck, Meike Hohwieler, Alexander Kleger, Matthias Meier
ABSTRACT

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.

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Anticorpo anti-Sox9, Chemicon®, from rabbit
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Anti-Rabbit-IgG - Atto 550 antibody produced in goat
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Anti-FLNB antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution