Testing susceptibility of patient-derived organoid cultures to therapies: Pharmacotyping

Richard Burkhart, Lindsey A. Baker, Hervé Tiriac

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages253-261
Number of pages9
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1787
ISSN (Print)1064-3745

Keywords

  • 3D culture
  • Drug testing
  • Organoids
  • Pharmacotyping
  • Tumor models

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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