Unit

UNIT 1.5 The Use of Fish-Derived Cell Lines for Investigation of Environmental Contaminants: An Update Following OECD's Fish Toxicity Testing Framework No. 171

  1. Vivian R. Dayeh1,
  2. Niels C. Bols1,
  3. Katrin Tanneberger2,
  4. Kristin Schirmer2,
  5. Lucy E. J. Lee3

Published Online: 1 MAY 2013

DOI: 10.1002/0471140856.tx0105s56

Current Protocols in Toxicology

Current Protocols in Toxicology

How to Cite

Dayeh, V. R., Bols, N. C., Tanneberger, K., Schirmer, K. and Lee, L. E. J. 2013. The Use of Fish-Derived Cell Lines for Investigation of Environmental Contaminants: An Update Following OECD's Fish Toxicity Testing Framework No. 171. Current Protocols in Toxicology. 56:1.5:1.5.1–1.5.20.

Author Information

  1. 1

    Department of Biology, University of Waterloo, Waterloo, Ontario, Canada

  2. 2

    Department of Environmental Toxicology, Eawag: Swiss Federal Institute of Aquatic Science and Technology, Dübendorf, Switzerland

  3. 3

    Department of Biology, University of the Fraser Valley, Abbotsford, British Columbia, Canada

Publication History

  1. Published Online: 1 MAY 2013

Abstract

Protocols for evaluating chemical toxicity at the cellular level using fish cell lines are described in this unit. Routine methodologies for growing salmonid cell lines, and using them in aquatic toxicology studies that support the mandate of the Organization for Economic Co-operation and Development (OECD) to reduce the use of whole animals in toxicity testing, are presented. Rapid, simple, cost-effective tests evaluating viability of cells with three indicator dyes per sample provides a broad overview of the sensitivity of cells to chemical contaminants. This fluorometric assay involves: (1) alamar blue for metabolic activity, (2) CFDA-AM for membrane integrity, and (3) neutral red for lysosomal function. These protocols are conveniently performed in semi-unison within the same multiwell plates and read at three different wavelengths. Detailed step-by-step descriptions of the assays, parameters to consider, troubleshooting, and guidelines for data interpretation are provided as essential tools for investigating environmental aquatic contaminants at the cellular level. Curr. Protoc. Toxicol. 56:1.5.1-1.5.20. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • fish cell lines;
  • in vitro toxicity;
  • fluorescent indicator dyes;
  • alamar Blue;
  • CFDA-AM;
  • neutral red;
  • toxicity test;
  • cell viability