Skip to Content
Merck
  • Modeling plasticity and dysplasia of pancreatic ductal organoids derived from human pluripotent stem cells.

Modeling plasticity and dysplasia of pancreatic ductal organoids derived from human pluripotent stem cells.

Cell stem cell (2021-04-30)
Markus Breunig, Jessica Merkle, Martin Wagner, Michael K Melzer, Thomas F E Barth, Thomas Engleitner, Johannes Krumm, Sandra Wiedenmann, Christian M Cohrs, Lukas Perkhofer, Gaurav Jain, Jana Krüger, Patrick C Hermann, Maximilian Schmid, Tamara Madácsy, Árpád Varga, Joscha Griger, Ninel Azoitei, Martin Müller, Oliver Wessely, Pamela G Robey, Sandra Heller, Zahra Dantes, Maximilian Reichert, Cagatay Günes, Christian Bolenz, Florian Kuhn, József Maléth, Stephan Speier, Stefan Liebau, Bence Sipos, Bernhard Kuster, Thomas Seufferlein, Roland Rad, Matthias Meier, Meike Hohwieler, Alexander Kleger
ABSTRACT

Personalized in vitro models for dysplasia and carcinogenesis in the pancreas have been constrained by insufficient differentiation of human pluripotent stem cells (hPSCs) into the exocrine pancreatic lineage. Here, we differentiate hPSCs into pancreatic duct-like organoids (PDLOs) with morphological, transcriptional, proteomic, and functional characteristics of human pancreatic ducts, further maturing upon transplantation into mice. PDLOs are generated from hPSCs inducibly expressing oncogenic GNAS, KRAS, or KRAS with genetic covariance of lost CDKN2A and from induced hPSCs derived from a McCune-Albright patient. Each oncogene causes a specific growth, structural, and molecular phenotype in vitro. While transplanted PDLOs with oncogenic KRAS alone form heterogenous dysplastic lesions or cancer, KRAS with CDKN2A loss develop dedifferentiated pancreatic ductal adenocarcinomas. In contrast, transplanted PDLOs with mutant GNAS lead to intraductal papillary mucinous neoplasia-like structures. Conclusively, PDLOs enable in vitro and in vivo studies of pancreatic plasticity, dysplasia, and cancer formation from a genetically defined background.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Dorsomorphin, ≥98% (HPLC)
Sigma-Aldrich
SANT-1, ≥98% (HPLC), powder
Sigma-Aldrich
3-Isobutyl-1-methylxanthine, ≥99% (HPLC), powder
Sigma-Aldrich
Monoclonal Anti-Glucagon antibody produced in mouse, clone K79bB10, ascites fluid
Sigma-Aldrich
Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-74, ascites fluid
Sigma-Aldrich
LDN193189 hydrochloride, ≥98% (HPLC)
Sigma-Aldrich
Dispase® II, protease
Sigma-Aldrich
Gastrin I human, ≥95% (HPLC), powder
Sigma-Aldrich
Carbonic Anhydrase from bovine erythrocytes, For use as a marker in SDS-PAGE
Sigma-Aldrich
Cholera Toxin B subunit, ≥95% (SDS-PAGE), lyophilized powder
Sigma-Aldrich
Anti-α-Amylase antibody produced in rabbit, fractionated antiserum, lyophilized powder
Sigma-Aldrich
N-Acetyl-L-cysteine, Sigma Grade, ≥99% (TLC), powder
Sigma-Aldrich
Anti-Vinculin antibody, Mouse monoclonal, clone hVIN-1, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Chymotrypsin Antibody, Human Pancreas, ascites fluid, Chemicon®
Sigma-Aldrich
Anti-Sox9 Antibody, Chemicon®, from rabbit