Skip to main content

Optimization of Cell Viability Assays for Drug Sensitivity Screens

  • Protocol
  • First Online:
Cell Viability Assays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2644))

  • 1904 Accesses

Abstract

During the preclinical stages of the drug discovery process, cell viability assays are fundamental tools for studying the phenotypic properties and overall health of cells following in vitro drug sensitivity screens. Therefore, it is important to optimize your viability assay of choice to obtain reproducible and replicable results, as well as use relevant drug response metrics (e.g., IC50, AUC, GR50, and GRmax) to identify candidate drugs for further evaluation in vivo. Herein, we used the resazurin reduction assay which is a quick, cost-effective, simple-to-use, and sensitive method for examining the phenotypic properties of cells. Using the MCF7 breast cancer cell line, we provide a detailed step-by-step protocol for optimizing drug sensitivity screens using the resazurin assay.

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

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 189.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
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

References

  1. Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, Minor L (2004) Cell viability assays. In: Sittampalam GS, Grossman A, Brimacombe K et al (eds) Assay guidance manual. Eli Lilly & Company and the National Center for Advancing Translational Sciences, Bethesda, pp 353–377

    Google Scholar 

  2. Stoddart MJ (2011) Cell viability assays: introduction. Methods Mol Biol 740:1–6. https://doi.org/10.1007/978-1-61779-108-6_1

    Article  CAS  PubMed  Google Scholar 

  3. Adan A, Kiraz Y, Baran Y (2016) Cell proliferation and cytotoxicity assays. Curr Pharm Biotechnol 17(14):1213–1221. https://doi.org/10.2174/1389201017666160808160513

    Article  CAS  PubMed  Google Scholar 

  4. Kamiloglu S, Sari G, Ozdal T, Capanoglu E (2020) Guidelines for cell viability assays. Food Front 1(3):332–349. https://doi.org/10.1002/fft2.44

    Article  Google Scholar 

  5. Aslantürk ÖS (2018) In vitro cytotoxicity and cell viability assays: principles, advantages, and disadvantages. In: Genotoxicity – a predictable risk to our actual world. IntechOpen, London, pp 13–30. https://doi.org/10.5772/intechopen.71923

    Chapter  Google Scholar 

  6. Präbst K, Engelhardt H, Ringgeler S, Hübner H (2017) Basic colorimetric proliferation assays: MTT, WST, and Resazurin. Methods Mol Biol 1601:1–17. https://doi.org/10.1007/978-1-4939-6960-9_1

    Article  CAS  PubMed  Google Scholar 

  7. Shokrzadeh M, Modanloo M (2017) An overview of the most common methods for assessing cell viability. J Res Med Dent Sci 5(2):33. https://doi.org/10.5455/jrmds.2017526

    Article  Google Scholar 

  8. Larsson P, Engqvist H, Biermann J, Werner Rönnerman E, Forssell-Aronsson E, Kovács A, Karlsson P, Helou K, Parris TZ (2020) Optimization of cell viability assays to improve replicability and reproducibility of cancer drug sensitivity screens. Sci Rep 10(1):5798. https://doi.org/10.1038/s41598-020-62848-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Ivanov DP, Parker TL, Walker DA, Alexander C, Ashford MB, Gellert PR, Garnett MC (2014) Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres. PLoS One 9(8):e103817. https://doi.org/10.1371/journal.pone.0103817

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ryu AH, Eckalbar WL, Kreimer A, Yosef N, Ahituv N (2017) Use antibiotics in cell culture with caution: genome-wide identification of antibiotic-induced changes in gene expression and regulation. Sci Rep 7(1):7533. https://doi.org/10.1038/s41598-017-07757-w

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Quast J-P (2022) Dose-response data analysis workflow. https://cran.r-project.org/web/packages/protti/vignettes/data_analysis_dose_response_workflow.html

  12. Hafner M, Niepel M, Subramanian K, Sorger PK (2017) Designing drug-response experiments and quantifying their results. Curr Protoc Chem Biol 9(2):96–116. https://doi.org/10.1002/cpch.19

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Hafner M, Niepel M, Chung M, Sorger PK (2016) Growth rate inhibition metrics correct for confounders in measuring sensitivity to cancer drugs. Nat Methods 13(6):521–527. https://doi.org/10.1038/nmeth.3853

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Niepel M, Hafner M, Chung M, Sorger PK (2017) Measuring cancer drug sensitivity and resistance in cultured cells. Curr Protoc Chem Biol 9(2):55–74. https://doi.org/10.1002/cpch.21

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Peter Larsson .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Larsson, P., Parris, T.Z. (2023). Optimization of Cell Viability Assays for Drug Sensitivity Screens. In: Friedrich, O., Gilbert, D.F. (eds) Cell Viability Assays. Methods in Molecular Biology, vol 2644. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3052-5_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3052-5_18

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3051-8

  • Online ISBN: 978-1-0716-3052-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics