Generation and Culture of Human Pancreatic Ductal Adenocarcinoma Organoids from Resected Tumor Specimens

  • Lindsey A. BakerEmail author
  • Hervé Tiriac
  • David A. TuvesonEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1882)


The recent development of human organoids as patient-specific models of pancreatic ductal adenocarcinoma (PDA) has helped set the stage for a new era of personalized medicine. Organoids can be generated from a resected PDA tumor in as little as 2–4 weeks, and are amenable to therapeutic screening as well as genetic and biochemical perturbation. Moreover, because these models promote the propagation of the neoplastic PDA cells at the expense of the stromal cells, transcriptome and genome-wide sequencing of organoids offers an unprecedented view of the genetic and expression changes occurring in the neoplastic cells of individual tumors. Here, we describe methods to generate PDA organoid cultures from resected human tumor specimens. We also describe how to propagate, cryopreserve, and thaw human PDA organoid cultures.

Key words

Pancreatic ductal adenocarcinoma Organoids Personalized medicine 3D culture Tumoroids Pancreatic cancer Patient-derived organoids 



We are grateful to Hans Clevers for an ongoing and productive collaboration to develop pancreatic cancer organoids, and to Candice Megan Young, Michelle Ma, and Hardik Patel for photography assistance and for thoughtful comments on this protocol. This work was supported through National Institute of Health (NIH) awards F32CA192904 (L.A.B.) and SWOG ITSC 5U10CA180944-04 (D.A.T., H.T.). In addition, 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); the NCI Cold Spring Harbor Laboratory (CSHL) Cancer Center Support Grant (5P30CA45508-29) and the Next Generation Sequencing and Microscopy 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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Cold Spring Harbor LaboratoryCold Spring HarborUSA
  2. 2.Lustgarten Foundation Pancreatic Cancer Research LaboratoryCold Spring HarborUSA

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