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

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Phenotypic Screening

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1787))

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.

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Correspondence to Richard A. Burkhart .

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Burkhart, R.A., Baker, L.A., Tiriac, H. (2018). Testing Susceptibility of Patient-Derived Organoid Cultures to Therapies: Pharmacotyping. In: Wagner, B. (eds) Phenotypic Screening. Methods in Molecular Biology, vol 1787. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7847-2_19

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  • DOI: https://doi.org/10.1007/978-1-4939-7847-2_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7846-5

  • Online ISBN: 978-1-4939-7847-2

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