Burkhart, R. A., Baker, L. A., Tiriac, H. (May 2018) Testing Susceptibility of Patient-Derived Organoid Cultures to Therapies: Pharmacotyping. Methods Mol Biol, 1787. pp. 253-261. ISSN 1064-3745
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.
Item Type: | Paper |
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Uncontrolled Keywords: | 3D culture Drug testing Organoids Pharmacotyping Tumor models |
Subjects: | Investigative techniques and equipment > cell culture > cancer organoids diseases & disorders > cancer > drugs and therapies |
CSHL Authors: | |
Communities: | CSHL labs > Tuveson lab |
Depositing User: | Matt Covey |
Date: | May 2018 |
Date Deposited: | 14 Jun 2018 21:20 |
Last Modified: | 14 Jun 2018 21:20 |
Related URLs: | |
URI: | https://repository.cshl.edu/id/eprint/36723 |
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