Cardiomyocyte H9c2 cells present a valuable alternative to fish lethal testing for azoxystrobin. (November 2015)

• Record Type: Journal Article
• Authors: Rodrigues, Elsa T.; Pardal, Miguel Â.; Laizé, Vincent; Cancela, M. Leonor; Oliveira, Paulo J.; Serafim, Teresa L.
• Is Part Of: Environmental pollution. Volume 206(2015)
• Page Start: 619
• Page End: 626
• Publication Date: 2015-11
• Subjects: Aquatic ecotoxicology -- Azoxystrobin -- Alternative assays -- Cell-based assays -- Cytotoxicity -- SRB assay; ΔΨm inner mitochondrial membrane potential -- AZX azoxystrobin -- CV coefficient of variation -- DMEM Dulbecco's modified Eagle's medium -- DMSO dimethyl sulphoxide -- FCCP p-trifluoromethoxy carbonyl cyanide phenyl hydrazine -- FBS foetal bovine serum -- IC50 median inhibition concentration -- KOW octanol-water partition coefficient -- LC50 median lethal concentration -- PBS phosphate buffered saline -- PMT photomultiplier tube -- SRB sulforhodamine B -- TMRE tetramethylrhodamine ethyl ester perchlorate -- Tris 2-amino-2-hydroxymethyl-propane-1, 3-diol -- ww wet weight; Pollution -- Periodicals; Pollution -- Environmental aspects -- Periodicals; Environmental Pollution -- Periodicals; Pollution -- Périodiques; Pollution -- Aspect de l'environnement -- Périodiques; Pollution -- Effets physiologiques -- Périodiques; Pollution; Pollution -- Environmental aspects; Periodicals; Electronic journals; 363.73
• DOI: 10.1016/j.envpol.2015.08.026
• Languages: English
• ISSNs: 0269-7491

Abstract

Abstract: The present study aims at identifying, among six mammalian and fish cell lines, a sensitive cell line whose in vitro median inhibitory concentration (IC50 ) better matches the in vivo short-term Sparus aurata median lethal concentration (LC50 ). IC50s and LC50 were assessed after exposure to the widely used fungicide azoxystrobin (AZX). Statistical results were relevant for most cell lines after 48 h of AZX exposure, being H9c2 the most sensitive cells, as well as the ones which provided the best prediction of fish toxicity, with a LC50, 96h /IC50, 48h = 0.581. H9c2 cell proliferation upon 72 h of AZX exposure revealed a LC50, 96h /IC50, 72h = 0.998. Therefore, identical absolute sensitivities were attained for both in vitro and in vivo assays. To conclude, the H9c2 cell-based assay is reliable and represents a suitable ethical alternative to conventional fish assays for AZX, and could be used to get valuable insights into the toxic effects of other pesticides. Highlights: Fish toxicity data are still considered standard information in ecotoxicology. Alternatives to animal testing have become an important topic of research. Cell-based assays are currently a promising in vitro alternative. Comparative studies to accelerate the validation of cell-based methods are required. H9c2 cell line proved to produce in vitro reliable toxicity results for azoxystrobin. Abstract : The application of cell-based assays for environmental toxicity studies would greatly reduce the number of fish needed for toxicity testing without any loss of reliability.