Testing Susceptibility of Patient-Derived Organoid Cultures to Therapies: Pharmacotyping

  • Richard A. Burkhart
  • Lindsey A. Baker
  • Hervé Tiriac
Part of the Methods in Molecular Biology book series (MIMB, volume 1787)


Increasingly, patient models of disease are being utilized to facilitate precision medicine approaches through molecular characterization or direct chemotherapeutic testing. Organoids, 3-dimensional (3D) cultures of neoplastic cells derived from primary tumor specimens, represent an ideal platform for these types of studies because benchtop protocols previously developed for 2-dimensional cell lines can be adapted for use. These protocols include directly testing the survival of these organoid cultures when exposed to clinically relevant chemotherapeutic agents, a process we have called pharmacotyping. In this protocol, established tumor-derived organoid cultures are dissociated into single cells, plated in a 3D gel matrix, and exposed to pharmacologic agents. While our protocol has been developed for use with patient-derived pancreatic ductal adenocarcinoma organoids, with minor modifications to the dissociation and medium conditions, this protocol could be adapted for use with a wide range of organoid cultures. We further describe our standard ATP-based assay to determine cellular survival. This protocol can be scaled for use in high-throughput assays.

Key words

3D culture Organoids Pharmacotyping Drug testing Tumor models 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Richard A. Burkhart
    • 1
    • 2
  • Lindsey A. Baker
    • 2
  • Hervé Tiriac
    • 2
  1. 1.Division of Hepatobiliary and Pancreatic Surgery, Department of SurgeryJohns Hopkins HospitalBaltimoreUSA
  2. 2.Lustgarten Foundation Pancreatic Cancer Research Laboratory, Cold Spring Harbor LaboratoryCold Spring HarborNew YorkUSA

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