Multiparametric luminescent cell viability assay in toxicology models: A critical evaluation

Journal of Pharmacological and Toxicological Methods
Nikolett SaliTamás Kőszegi

Abstract

In cellular viability assays the sole determination of a single parameter might not give precise information on the extent of toxicity. In our study we worked out a multiparametric microplate assay based on bioluminescent ATP quantification, esterase activity-related fluorescence, nucleic acid staining and total intracellular protein measurement from the same sample in MDCK and HepG2 tissue cultures. Dose-response analyses were done after ATP depletion by metabolic poisons (NaF, NaN3) and by ochratoxin A (OTA) mycotoxin treatments. A novel perchloric acid fixation/extraction technique was applied in order to obtain intracellular ATP levels, esterase activity, DNA content and protein data simultaneously. Esterase activity was assessed by a fluorogenic staining. Estimation of cell number was done by DAPI fluorescence. Our results were expressed as ATP/protein, calcein fluorescence/ATP, calcein fluorescence/protein and ATP/DAPI ratios. Apoptosis/necrosis rates were measured by Annexin V-propidium iodide and 7-aminoactinomycin D flow cytometric assays and effects of OTA on actin cytoskeleton were also studied by using labeled phalloidin for visualization of actin. We could verify that the esterase assay was not an energy driven (tr...Continue Reading

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Citations

Feb 15, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Rita CsepregiMiklós Poór
Jan 26, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sourav DasTamás Kőszegi
Sep 13, 2018·International Journal of Molecular Sciences·Rita CsepregiTamás Kőszegi
Nov 15, 2018·Mycotoxin Research·Zelma FaisalMiklós Poór
May 28, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sourav DasAleksandar Széchenyi
Jul 1, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Rita CsepregiTamás Kőszegi

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