The comet assay applied to HepG2 liver spheroids.
Cell Culture Techniques
Cell Survival
/ drug effects
Chlorpromazine
/ toxicity
Colchicine
/ toxicity
Comet Assay
/ methods
DNA Damage
DNA, Neoplasm
/ drug effects
Guanine
/ analogs & derivatives
Hep G2 Cells
/ drug effects
Humans
Hydrogen Peroxide
/ toxicity
Laboratory Proficiency Testing
Methyl Methanesulfonate
/ toxicity
Sensitivity and Specificity
Spheroids, Cellular
/ drug effects
2D and 3D culture
Advanced in vitro model
Comet assay
Genotoxicity
Liver spheroids
Journal
Mutation research. Genetic toxicology and environmental mutagenesis
ISSN: 1879-3592
Titre abrégé: Mutat Res Genet Toxicol Environ Mutagen
Pays: Netherlands
ID NLM: 101632149
Informations de publication
Date de publication:
Sep 2019
Sep 2019
Historique:
received:
21
11
2018
revised:
06
03
2019
accepted:
15
03
2019
entrez:
29
9
2019
pubmed:
29
9
2019
medline:
24
3
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31561895
pii: S1383-5718(18)30424-8
doi: 10.1016/j.mrgentox.2019.03.006
pii:
doi:
Substances chimiques
DNA, Neoplasm
0
8-hydroxyguanine
5614-64-2
Guanine
5Z93L87A1R
Methyl Methanesulfonate
AT5C31J09G
Hydrogen Peroxide
BBX060AN9V
Colchicine
SML2Y3J35T
Chlorpromazine
U42B7VYA4P
Types de publication
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
403033Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.