Advertisement

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

  • Richard A. Burkhart
  • Lindsey A. Baker
  • Hervé Tiriac
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

3D culture Organoids Pharmacotyping Drug testing Tumor models 

References

  1. 1.
    Howlader N, Noone A, Krapcho M et al. (2016) SEER cancer statistics review, 1975–2013. National Cancer Institute. http://seer.cancer.gov/csr/1975_2013/. Accessed July 2016
  2. 2.
    DeMatteo RP, Ballman KV, Antonescu CR, Corless C, Kolesnikova V, von Mehren M et al (2013) Long-term results of adjuvant imatinib mesylate in localized, high-risk, primary gastrointestinal stromal tumor: ACOSOG Z9000 (alliance) intergroup phase 2 trial. Ann Surg 258:422–429CrossRefGoogle Scholar
  3. 3.
    Alexandrov LB, Nik-Zainal S, Wedge DC, Aparicio SA, Behjati S, Biankin AV et al (2013) Signatures of mutational processes in human cancer. Nature 500:415–421CrossRefGoogle Scholar
  4. 4.
    Lowery MA, Jordan EJ, Basturk O, Ptashkin RN, Zehir A, Berger MF et al (2017) Real time genomic profiling of pancreatic ductal adenocarcinoma: potential actionability and correlation with clinical phenotype. Clin Cancer Res 23:6094–6100. pii: clincanres.0899.2017CrossRefGoogle Scholar
  5. 5.
    Witkiewicz AK, Balaji U, Eslinger C, McMillan E, Conway W, Posner B et al (2016) Integrated patient-derived models delineate individualized therapeutic vulnerabilities of pancreatic cancer. Cell Rep 16:2017–2031CrossRefGoogle Scholar
  6. 6.
    Boj SF, Hwang CI, Baker LA, Chio II, Engle DD, Corbo V et al (2015) Organoid models of human and mouse ductal pancreatic cancer. Cell 160:324–338CrossRefGoogle Scholar

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

Personalised recommendations