Abstract
Pancreatic ductal adenocarcinoma (PDA) is a lethal malignancy that is refractory to all current therapies. Research into the mechanisms driving this cancer is the key to developing better diagnostic and treatment options which are urgently needed in the clinic. Genetically engineered mouse models of PDA have been valuable research tools, enabling studies of all stages of PDA progression. However, these models are difficult and time-consuming to breed, and engineering further mutations into these models requires additional time. Recently, organoid cultures of PDA have emerged as alternative models for this disease. Organoids can be rapidly generated from mouse models of PDA and enable genetic and biochemical perturbation of all stages of PDA progression. Here, we describe the generation and propagation of organoid models from PDA tumors and metastases harvested from genetically engineered mouse models.
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Acknowledgments
We are grateful to Hans Clevers for an ongoing and productive collaboration to develop pancreatic cancer organoids, to Mona S. Spector and Sylvia F. Boj for initial development of the murine PDA organoid methods, and to Dannielle D. Engle, Youngkyu Park, and Melissa Yao for their thoughtful comments on this protocol. L.A.B. was supported through the National Institute of Health (NIH, F32CA192904). D.A.T. was supported by NIH awards 1R01CA190092-04, 1R01CA188134-01, 5P20CA192996-03, 5P50CA101955-07, 5U01CA168409-5, 1U10CA180944-04, 1U01CA210240-01A1, and P20CA19299402; D.O.D. award W81XWH-13-PRCRP-IA; a gift from the Simons Foundation (552716); the STARR Cancer Consortium (I7-A718); SWOG ITSC 5U10CA180944-04; the NCI Cold Spring Harbor Laboratory (CSHL) Cancer Center Support Grant (5P30CA45508-29) and the Animal, Genetic Engineering, Microscopy, and Next Generation Sequencing Shared Resources; the V Foundation; the Thompson Family Foundation; Stand Up to Cancer/KWF (SU2C-AACR-PS09); the Precision Medicine Research Associates; the CSHL Association; and by the Lustgarten Foundation, where D.A.T. is a distinguished scholar and Director of the Lustgarten Foundation-designated Laboratory of Pancreatic Cancer Research and the Director of the CSHL Cancer Center.
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Baker, L.A., Tuveson, D.A. (2019). Generation and Culture of Tumor and Metastatic Organoids from Murine Models of Pancreatic Ductal Adenocarcinoma. In: Su, G. (eds) Pancreatic Cancer. Methods in Molecular Biology, vol 1882. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8879-2_10
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DOI: https://doi.org/10.1007/978-1-4939-8879-2_10
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