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Repeatability and reproducibility of the RTgill-W1 cell line assay for predicting fish acute toxicity.

Predicting fish acute toxicity of chemicals in vitro is an attractive alternative method to the conventional approach using juvenile and adult fish. The rainbow trout (Oncorhynchus mykiss) cell line assay with RTgill-W1 cells has been designed for this purpose. It quantifies cell viability using fluorescent measurements for metabolic activity, cell- and lysosomal- membrane integrity on the same set of cells. Results from over 70 organic chemicals attest to the high predictive capacity of this test. We here report on the repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of the RTgill-W1 cell line assay in a round-robin study focusing on six test chemicals involving six laboratories from the industrial and academic sector. All participating laboratories were able to establish the assay according to pre-set quality criteria even though, apart from the lead laboratory, none had previously worked with the RTgill-W1 cell line. Concentration-response modelling, based on either nominal or geometric mean-derived measured concentrations, yielded effect concentrations (EC50) that spanned approximately four orders of magnitude over the chemical range, covering all fish acute toxicity categories. Coefficients of variation for intra and inter-laboratory variability for the average of the three fluorescent cell viability measurements were 15.5% and 30.8%, respectively, which is comparable to other fish-derived, small scale bioassays. This study therefore underlines the robustness of the RTgill-W1 cell line assay and its accurate performance when carried out by operators in different laboratory settings.

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