Evaluation of alamar blue reduction for the in vitro assay of hepatocyte toxicity

Toxicology in Vitro : an International Journal Published in Association with BIBRA
M R SlaughterP J O'Brien

Abstract

Alamar Blue (AB) reduction is a promising new in vitro assay which is simple to conduct and amenable to repeated measurements and high-throughput screening; however, evaluation with hepatocytes has not been reported. Accordingly, we compared AB reduction with established markers of hepatocyte viability and cell density. Primary rat hepatocytes were allowed to adhere to collagen-coated 96-well plates, then exposed for 16 hours to culture medium, 0.7% dimethyl sulfoxide (DMSO) in medium, 500 mum CCl(4), 500 mum eugenol or 15 or 150 mum of a novel substituted indoline (the latter three in medium with 0.7% DMSO; medium also contained hydrocortisone during exposure period). Using a spectrophotometric plate reader, AB reduction was compared with lactate dehydrogenase release (LDH) release, neutral red (NR) uptake, total protein (TP) and cell seed density. AB reduction showed a linear relationship and good correlation with NR uptake, LDH release, TP and cell density. AB assay precision varied with cell density, but was similar to other assays in cytotoxicity screening. Good correlation with cell density indicates AB to have the potential for assessment of hepatocyte proliferation. From the results reported here, we recommend further e...Continue Reading

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Citations

Apr 14, 2005·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Willem G E J SchoonenEric Débiton
May 30, 2001·Toxicology in Vitro : an International Journal Published in Association with BIBRA·R J Gonzalez, J B Tarloff
Apr 20, 2004·Assay and Drug Development Technologies·Yuansheng Yang, R Robert Balcarcel
Jan 20, 2012·The Journal of Antimicrobial Chemotherapy·Manal J NattoHarry P de Koning
Jul 24, 2014·Pesticide Biochemistry and Physiology·G TajuA S Sahul Hameed
Nov 3, 2009·Toxicology Letters·Isabelle SeverinMarie-Christine Chagnon
Sep 3, 2010·Journal of Applied Toxicology : JAT·Saura C SahuPaddy L Wiesenfeld
Jul 11, 2012·Journal of Applied Toxicology : JAT·Saura C SahuRobert L Sprando
Feb 14, 2014·Journal of Applied Toxicology : JAT·Saura C SahuRobert L Sprando
Dec 22, 2007·Journal of Biomedical Materials Research. Part B, Applied Biomaterials·Martin RosentrittGerhard Handel
Apr 25, 2015·Drug Discovery Today·David J BraydenJeremy C Simpson
Mar 12, 2016·PLoS Neglected Tropical Diseases·Britta StadelmannAndrew Hemphill
Sep 29, 2011·Journal of Applied Toxicology : JAT·Saura C SahuRobert L Sprando
Sep 11, 2004·Assay and Drug Development Technologies·Yuansheng Yang, R Robert Balcarcel
Feb 2, 2010·Journal of Biomechanics·C FriasA T Marques

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