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The comet assay applied to HepG2 liver spheroids

Elje, Elisabeth ; Hesler, Michelle ; Rundén-Pran, Elise ; Mann, Pascal ; Mariussen, Espen ; Wagner, Sylvia ; Dusinska, Maria ; Kohl, Yvonne

Mutation research, 2019-09, Vol.845, p.403033, Article 403033 [Periódico revisado por pares]

Netherlands: Elsevier B.V

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The comet assay applied to HepG2 liver spheroids
Autor: Elje, Elisabeth ; Hesler, Michelle ; Rundén-Pran, Elise ; Mann, Pascal ; Mariussen, Espen ; Wagner, Sylvia ; Dusinska, Maria ; Kohl, Yvonne
Assuntos: 2D and 3D culture ; Advanced in vitro model ; Cell Culture Techniques ; Cell Survival - drug effects ; Chlorpromazine - toxicity ; Colchicine - toxicity ; Comet assay ; Comet Assay - methods ; Comet Assay - standards ; DNA Damage ; DNA, Neoplasm - drug effects ; DNA, Neoplasm - genetics ; Genotoxicity ; Guanine - analogs & derivatives ; Guanine - analysis ; Hep G2 Cells - drug effects ; Humans ; Hydrogen Peroxide - toxicity ; Laboratory Proficiency Testing ; Liver spheroids ; Methyl Methanesulfonate - toxicity ; Sensitivity and Specificity ; Spheroids, Cellular - drug effects
É parte de: Mutation research, 2019-09, Vol.845, p.403033, Article 403033
Descrição: •HepG2 liver spheroids preparation was standardized in independent laboratories.•Comet assay was applied successfully to HepG2 liver spheroids for the first time.•A difference in sensitivity was found comparing the response of 2D and 3D cultures. In accordance with the 3 Rs to reduce in vivo testing, more advanced in vitro models, moving from 2D monolayer to 3D cultures, should be developed for prediction of human toxicity of industrial chemicals and environmental pollutants. In this study we compared cytotoxic and genotoxic responses induced by chemicals in 2D and 3D spheroidal cultures of the human liver cancer cell line HepG2. HepG2 spheroids were prepared by hanging drop technology. Both 3D spheroids and 2D monolayer cultures were exposed to different chemicals (colchicine, chlorpromazine hydrochloride or methyl methanesulfonate) for geno- and cytotoxicity studies. Cytotoxicity was investigated by alamarBlue assay, flow cytometry and confocal imaging. DNA damage was investigated by the comet assay with and without Fpg enzyme for detection of DNA strand breaks and oxidized or alkylated base lesions. The results from the cyto- and genotoxicity tests showed differences in sensitivity comparing the 2D and 3D HepG2 models. This study shows that human 3D spheroidal hepatocellular cultures can be successfully applied for genotoxicity testing by the comet assay and represent a promising advanced in vitro model for toxicity testing.
Editor: Netherlands: Elsevier B.V
Idioma: Inglês

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The comet assay applied to HepG2 liver spheroids

Elje, Elisabeth ; Hesler, Michelle ; Rundén-Pran, Elise ; Mann, Pascal ; Mariussen, Espen ; Wagner, Sylvia ; Dusinska, Maria ; Kohl, Yvonne

Netherlands: Elsevier B.V

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