Viability assay with fish gill cell line to assess acute toxicity
Commonly used acronym: RTgill-W1 cell line assay
Scope of the method
- Translational - Applied Research
- In vitro - Ex vivo
- Animal derived cells / tissues / organs
- cell viability test
- fish gill cell line
- cell metabolic activity
- lysosomal membrane integrity
- cell membrane integrity
- fish acute toxicity
- chemical exposure
The rainbow trout gill cell line assay quantifies cell viability using fluorescent measurements for metabolic activity (Alamar Blue, AB), cell membrane integrity (5-CarboxyFluorescein DiAcetate AcetoxyMethyl ester, CFDA-AM) and lysosomal membrane integrity (Neutral Red, NR). Chemicals are added to confluent RTgill-W1 cell monolayers in 24-well plates with L-15/ex medium (a simplified version of L-15 cell culture medium without serum). Cells are incubated for 24 hours in the incubator (19°C, normal atmosphere, in the dark). At the end of the exposure, cell viability measurements are performed with 3 fluorescent indicator dyes on the same set of exposed cells.
- Laminar flow ;
- Incubator (room temperature, no CO2) ;
- Microplate reader for fluorescence detection.
- History of use
- Internally validated
- Published in peer reviewed journal
Pros, cons & Future potential
- Cell line model with limited requirements ;
- Robust assay: repeatability and reproducibility is shown through inter- and intralaboratory studies ;
- Alternative model to predict fish acute toxicity.
Exposure of chemicals (bioavailability).
References, associated documents and other information
Fischer, M ; Belanger, SE ; Berckmans, P ; Bernhard, MJ ; Blaha, L ; Schmid, DEC ; Dyer, SD ; Haupt, T ; Hermens, JLM ; Hultman, MT ; Laue, H ; Lillicrap, A ; Minarikova, M ; Natsch, A ; Novak, J ; Sinnige, TL ; Tollefsen, KE ; von Niederhausern, V ; Witters, H ; Zupanic, A ; & K. Schirmer (2019). Repeatability and reproducibility of the RTgill-W1 cell line assay for predicting fish acute toxicity. Toxicological Sciences, 169 (2), 353-3640.
Tanneberger, K., Knoebel, M., Busser, F. J. M., Sinnige, T. L.,Hermens, J. L. M., and Schirmer, K. (2013). Predicting fish acute toxicity using a fish gill cell line-based toxicity assay. Environ. Sci. Technol. 47, 1110–1119.
ISO 21115:2019. Water quality — Determination of acute toxicity of water samples and chemicals to a fish gill cell line (RTgill-W1).