Title :

DMSO-free highly differentiated HepaRG spheroids for chronic toxicity, liver functions and genotoxicity studies.

Author(s) :

Rose, S. [Auteur]
Institut de recherche en santé, environnement et travail [Irset]
Cuvellier, M. [Auteur]
Ezan, F. [Auteur]
Carteret, J. [Auteur]
Bruyère, A. [Auteur]
Legagneux, V. [Auteur]
Nesslany, Fabrice [Auteur]
IMPact de l'Environnement Chimique sur la Santé humaine (IMPECS) - ULR 4483
Baffet, G. [Auteur]
Langouët, S. [Auteur]

Journal title :

Archives of Toxicology

Abbreviated title :

Arch Toxicol

Publication date :

2021-11-20

ISSN :

1432-0738

English keyword(s) :

HepaRG cells
3D culture model
Long-term differentiation
Genotoxicity
Human liver injury
Toxicity screening

HAL domain(s) :

Sciences du Vivant [q-bio]

English abstract : [en]

The liver is essential in the elimination of environmental and food contaminants. Given the interspecies differences between rodents and humans, the development of relevant in vitro human models is crucial to investigate ...
Show more >The liver is essential in the elimination of environmental and food contaminants. Given the interspecies differences between rodents and humans, the development of relevant in vitro human models is crucial to investigate liver functions and toxicity in cells that better reflect pathophysiological processes. Classically, the differentiation of the hepatic HepaRG cell line requires high concentration of dimethyl sulfoxide (DMSO), which restricts its usefulness for drug-metabolism studies. Herein, we describe undifferentiated HepaRG cells embedded in a collagen matrix in DMSO-free conditions that rapidly organize into polarized hollow spheroids of differentiated hepatocyte-like cells (Hepoid-HepaRG). Our conditions allow concomitant proliferation with high levels of liver-specific functions and xenobiotic metabolism enzymes expression and activities after a few days of culture and for at least 4 weeks. By studying the toxicity of well-known injury-inducing drugs by treating cells with 1- to 100-fold of their plasmatic concentrations, we showed appropriate responses and demonstrate the sensitivity to drugs known to induce various degrees of liver injury. Our results also demonstrated that the model is well suited to estimate cholestasis and steatosis effects of drugs following chronic treatment. Additionally, DNA alterations caused by four genotoxic compounds (Aflatoxin B1 (AFB1), Benzo[a]Pyrene (B[a]P), Cyclophosphamide (CPA) and Methyl methanesulfonate (MMS)) were quantified in a dose-dependent manner by the comet and micronucleus assays. Their genotoxic effects were significantly increased after either an acute 24 h treatment (AFB1: 1.5–6 μM, CPA: 2.5–10 μM, B[a]P: 12.5–50 μM, MMS: 90–450 μM) or after a 14-day treatment at much lower concentrations (AFB1: 0.05–0.2 μM, CPA: 0.125–0.5 μM, B[a]P: 0.125–0.5 μM) representative to human exposure. Altogether, the DMSO-free 3D culture of Hepoid-HepaRG provides highly differentiated and proliferating cells relevant for various toxicological in vitro assays, especially for drug-preclinical studies and environmental chemicals risk assessment.Show less >

Language :

Anglais

Audience :

Internationale

Popular science :

Non

Administrative institution(s) :

Université de Lille
CHU Lille
Institut Pasteur de Lille

Collections :

• IMPact de l'Environnement Chimique sur la Santé humaine (IMPECS) - ULR 4483

Submission date :

2023-10-20T06:13:48Z
2024-02-23T14:32:54Z