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Cross-Species Extrapolation Using a Simplified In Vitro Tissue Explant Assay in Fish

  • Bryanna Eisner
  • Jon Doering
  • Shawn Beitel
  • Markus Hecker
Protocol
Part of the Methods in Pharmacology and Toxicology book series

Abstract

Legislation in North America and Europe, such as the Canadian Environmental Protection Act (CEPA), the US Safe Drinking Water Act (SDWA), and the European Registration, Evaluation, and Authorization of Chemicals (REACH) program, mandates the assessment of risks of chemicals to the environment and human health. Current testing strategies within these pieces of legislation rely on extensive animal testing and on extrapolation from standard laboratory model species to native species of relevance in local ecosystems or humans. In addition to the huge costs and large numbers of animals needed, it becomes apparent that these approaches are often not adequately predictive and, thus, protective of organisms of interest. Furthermore, there are significant ethical concerns due to the large numbers of live animals these testing efforts require. For this reason, there is a need for alternative in vitro testing approaches that enable extrapolating to in vivo results and address the lack of toxicity data for the diversity of organisms. One such in vitro method is the simplified tissue explant assay, in which tissues are excised from individuals and maintained and exposed in a culture over a few hours to several days. Tissue explants have many advantages over classic cell line-based assays, such as maintenance of paracrine cell to cell interactions, and have also shown to be less variable in some cases. Most importantly, tissue explants maintain species-specific properties that enable more realistic characterization of contaminant effects in organisms of interest. Numerous different tissues, such as liver, gonads, and lung/gill, from several taxonomic groups, including species of mammals, birds, amphibians, and fish, have been successfully cultured and exposed to contaminants. After the exposure, various biochemical endpoints as well as overall cell viability can be assessed. In this chapter, we discuss the advantages of using tissue explant assays over stable cell lines, with a focus on fish species, which has been the most commonly used taxonomic group for this purpose to date. We then outline the steps involved in conducting exposure experiments with species-specific tissue explants and provide detailed guidelines to assist in successfully carrying out and interpreting tissue explant experiments.

Keywords

In vitro Tissue explants Alternative testing methods Cross-species extrapolation High-throughput 

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Copyright information

© Springer Science+Business Media, LLC 2019

Authors and Affiliations

  • Bryanna Eisner
    • 1
  • Jon Doering
    • 1
  • Shawn Beitel
    • 1
  • Markus Hecker
    • 1
  1. 1.Toxicology CentreUniversity of SaskatchewanSaskatoonCanada

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