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High-Throughput Assessment of Metabolism-Induced Toxicity of Compounds on a 384-Pillar Plate

  • Soo-Yeon Kang
  • Kyeong-Nam Yu
  • Pranav Joshi
  • Moo-Yeal LeeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2089)

Abstract

A variety of oxidative and conjugative enzymes are involved in the metabolism of compounds including drugs, which can be converted into toxic metabolites by Phase I drug-metabolizing enzymes (DMEs), such as the cytochromes P450 (CYP450s), and/or detoxified by Phase II DMEs, such as UDP-glucuronosyltransferases (UGTs), sulfotransferases (SULTs), and glutathione S-transferases (GSTs). Traditionally, primary hepatocytes containing a complete set of DMEs have been widely used as a gold standard to assess metabolism-induced compound toxicity. However, primary hepatocytes are expensive, have high donor variability in expression levels of DMEs, and rapidly lose liver-specific functions when the cells are maintained under standard in vitro cell culture conditions over time. To address this issue and rapidly profile metabolism-induced drug toxicity, we have developed a 384-pillar plate, which is complementary to conventional 384-well plates. In this chapter, we provide step-by-step procedures for three-dimensional (3D) cell printing on the 384-pillar plate coupled with DMEs and compounds in the 384-well plate for high-throughput assessment of metabolism-induced toxicity.

Key words

Metabolism-induced toxicity Drug-metabolizing enzymes 384-Pillar plate 3D cell culture High-throughput screening (HTS) 

Notes

Acknowledgments

This work was partially supported by the US Environmental Protection Agency (US EPA Transform Tox Testing Challenge), Medical & Bio Device (MBD) Korea, the Cleveland State University (Faculty Innovation Fund), and the National Institutes of Health (NIEHS R01ES025779).

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Copyright information

© Springer Science+Business Media, LLC 2020

Authors and Affiliations

  • Soo-Yeon Kang
    • 1
  • Kyeong-Nam Yu
    • 1
  • Pranav Joshi
    • 1
  • Moo-Yeal Lee
    • 1
    Email author
  1. 1.Department of Chemical and Biomedical EngineeringCleveland State UniversityClevelandUSA

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