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High-Throughput Cell Toxicity Assays

  • David MurrayEmail author
  • Lisa McWilliams
  • Mark Wigglesworth
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1439)

Abstract

Understanding compound-driven cell toxicity is vitally important for all drug discovery approaches. With high-throughput screening (HTS) being the key strategy to find hit and lead compounds for drug discovery projects in the pharmaceutical industry [1], an understanding of the cell toxicity profile of hit molecules from HTS activities is fundamentally important. Recently, there has been a resurgence of interest in phenotypic drug discovery and these cell-based assays are now being run in HTS labs on ever increasing numbers of compounds. As the use of cell assays increases the ability to measure toxicity of compounds on a large scale becomes increasingly important to ensure that false hits are not progressed and that compounds do not carry forward a toxic liability that may cause them to fail at later stages of a project. Here we describe methods employed in the AstraZeneca HTS laboratory to carry out very large scale cell toxicity screening.

Key words

Toxicity Cell-based assays Phenotypic High-throughput screening HTS Resazurin Apoptosis 

References

  1. 1.
    Macarron R et al (2011) Impact of high-throughput screening in biomedical research. Nat Rev Drug Discov 10:188–195CrossRefGoogle Scholar
  2. 2.
    Mayr LM, Bojanic D (2009) Novel trends in high-throughput screening. Curr Opin Pharmacol 9:580–588Google Scholar
  3. 3.
    Cook D et al (2014) Lessons learned from the fate of AstraZeneca’s drug pipeline: a five-dimensional framework. Nat Rev Drug Discov 13:419–431CrossRefGoogle Scholar
  4. 4.
    O’Brien J et al (2000) Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem 267:5421–5426CrossRefGoogle Scholar
  5. 5.
    Hamid R et al (2004) Comparison of alamar blue and MTT assays for high through-put screening. Toxicol In Vitro 18(2004):703–710CrossRefGoogle Scholar
  6. 6.
    Nakayama GR et al (1997) Assessment of the Alamar Blue assay for cellular growth and viability in vitro. J Immunol Methods 204:205–208CrossRefGoogle Scholar
  7. 7.
    Slater K (2001) Cytotoxicity tests for high-throughput drug discovery. Curr Opin Biotechnol 12:70–74CrossRefGoogle Scholar
  8. 8.
    Weyermann J et al (2005) A practical note on the use of cytotoxicity assays. Int J Pharm 288:369–376CrossRefGoogle Scholar
  9. 9.
    Promega Corporation. CellTox™ Green Cytotoxicity Assay. https://www.promega.com/products/cell-health-and-metabolism/cytotoxicity-assays/real-time-cytotoxicity-assay/celltox-green-cytotoxicity-assay/?activeTab= 0. Accessed 1 July 2015Google Scholar
  10. 10.
    McDougal M et al (2010) Nucleic acid binding dyes and uses thereof. US Patent 2010/0233710 A1Google Scholar
  11. 11.
    Korzeniewski C, Callewaert DM (1983) An enzyme-release assay for natural cytotoxicity. J Immunol Methods 64:313–320CrossRefGoogle Scholar
  12. 12.
    Lobner D (2000) Comparison of the LDH and MTT assays for quantifying cell death: validity for neuronal apoptosis? J Neurosci Methods 96:147–152CrossRefGoogle Scholar
  13. 13.
    Fotakis G, Timbrell JA (2006) In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett 160:171–177CrossRefGoogle Scholar
  14. 14.
    Luu YK et al (2012) Profiling of toxicity and identification of distinct apoptosis profiles using a 384-well high-throughput flow cytometry screening platform. J Biomol Screen 17:806–812CrossRefGoogle Scholar
  15. 15.
    Griffith D et al (2012) Implementation and development of an automated, ultra-high-capacity, acoustic, flexible dispensing platform for assay-ready plate delivery. J Lab Autom 17(5):348–358CrossRefGoogle Scholar
  16. 16.
    Zhang J-H et al (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 4:67–73CrossRefGoogle Scholar
  17. 17.
    Lundholt BK et al (2003) A simple technique for reducing edge effect in cell-based assays. J Biomol Screen 8:566–570CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • David Murray
    • 1
    Email author
  • Lisa McWilliams
    • 1
  • Mark Wigglesworth
    • 1
  1. 1.Discovery Sciences, Innovative MedicinesAstraZenecaMacclesfieldUK

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