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Development of a rainbow trout intestinal epithelial cell line and its response to lipopolysaccharide

2010; Wiley; Volume: 17; Issue: 2 Linguagem: Inglês

10.1111/j.1365-2095.2010.00757.x

1365-2095

Ayumi Kawano, Christian Haiduk, Kristin Schirmer, Robert Hanner, Lucy E. J. Lee, Brian Dixon, Niels C. Bols,

Animal Genetics and Reproduction

Aquaculture NutritionVolume 17, Issue 2 p. e241-e252 Development of a rainbow trout intestinal epithelial cell line and its response to lipopolysaccharide A. KAWANO, A. KAWANO Department of Biology, University of Waterloo, Waterloo, Ontario, CanadaSearch for more papers by this authorC. HAIDUK, C. HAIDUK UFZ, Helmholtz Centre for Environmental Research, Leipzig, GermanySearch for more papers by this authorK. SCHIRMER, K. SCHIRMER Eawag–Swiss Federal Institute of Aquatic Science and Technology, Dübendorf, SwitzerlandSearch for more papers by this authorR. HANNER, R. HANNER Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario, CanadaSearch for more papers by this authorL.E.J. LEE, L.E.J. LEE Department of Biology, Wilfrid Laurier University, Waterloo, Ontario, CanadaSearch for more papers by this authorB. DIXON, B. DIXON Department of Biology, University of Waterloo, Waterloo, Ontario, CanadaSearch for more papers by this authorN.C. BOLS, N.C. BOLS Department of Biology, University of Waterloo, Waterloo, Ontario, CanadaSearch for more papers by this author A. KAWANO, A. KAWANO Department of Biology, University of Waterloo, Waterloo, Ontario, CanadaSearch for more papers by this authorC. HAIDUK, C. HAIDUK UFZ, Helmholtz Centre for Environmental Research, Leipzig, GermanySearch for more papers by this authorK. SCHIRMER, K. SCHIRMER Eawag–Swiss Federal Institute of Aquatic Science and Technology, Dübendorf, SwitzerlandSearch for more papers by this authorR. HANNER, R. HANNER Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario, CanadaSearch for more papers by this authorL.E.J. LEE, L.E.J. LEE Department of Biology, Wilfrid Laurier University, Waterloo, Ontario, CanadaSearch for more papers by this authorB. DIXON, B. DIXON Department of Biology, University of Waterloo, Waterloo, Ontario, CanadaSearch for more papers by this authorN.C. BOLS, N.C. BOLS Department of Biology, University of Waterloo, Waterloo, Ontario, CanadaSearch for more papers by this author First published: 10 March 2011 https://doi.org/10.1111/j.1365-2095.2010.00757.xCitations: 94 Correspondence: Niels Bols, University of Waterloo, 200 University Ave, Waterloo, Ontario, Canada N2L 3G1. E-mail:[email protected] Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Abstract A cell line, RTgutGC, was developed from the intestine of Oncorhynchus mykiss. RTgutGC has an epithelial-like shape, been passaged over 100 times, and cryopreserved successfully. A rainbow trout origin was confirmed by sequencing a 652 bp region of the mitochondrial cytochrome c oxidase I gene. RTgutGC is grown routinely in Leibovitz’s L15 without glutamine supplemented with 10% fetal bovine serum (FBS). Cell viability was evaluated with Alamar blue (AB) for metabolic activity and carboxyfluorescein diacetate acetoxymethyl ester (CFDA AM) for membrane integrity. Viability was unchanged by lipopolysaccharide (LPS) for cultures in FBS. For cultures at low cell densities in L15 without FBS or glutamine, cell viability declined in a LPS dose-dependent manner, allowing calculation of the concentration causing a 50% decline in viability (EC50). When glutamine was present, the EC50 was increased for both AB and CFDA AM. As the cell density increased, LPS became much less cytotoxic and no EC50 could be calculated for very confluent cultures. 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