Skip to main content

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

  • Protocol
  • First Online:
Phenotypic Screening

Part of the book series: Methods in Molecular Biology ((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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others


  1. Howlader N, Noone A, Krapcho M et al. (2016) SEER cancer statistics review, 1975–2013. National Cancer Institute. Accessed July 2016

  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 258:422–429

    Article  Google Scholar 

  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 500:415–421

    Article  CAS  Google Scholar 

  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.0899.2017

    Article  CAS  Google Scholar 

  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 16:2017–2031

    Article  CAS  Google Scholar 

  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 160:324–338

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to Richard A. Burkhart .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

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

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

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.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7846-5

  • Online ISBN: 978-1-4939-7847-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics