title: Three-dimensional HepaRG model as an attractive tool for toxicity testing.
abstract: The culture of HepaRG cells as three dimensional (3D) structures in the spinner-bioreactor may represent added value as a hepatic system for toxicological purposes. The use of a cost-effective commercially available bioreactor, which is compatible with high-throughput cell analysis, constitutes an attractive approach for routine use in the drug testing industry. In order to assess specific aspects of the biotransformation capacity of the bioreactor-based HepaRG system, the induction of CYP450 enzymes (i.e., CYP1A2, 2B6, 2C9, and 3A4) and the activity of the phase II enzyme, uridine diphosphate glucuronoltransferase (UGT), were tested. The long-term functionality of the system was demonstrated by 7-week stable profiles of albumin secretion, CYP3A4 induction, and UGT activities. Immunofluorescence-based staining showed formation of tissue-like arrangements including bile canaliculi-like structures and polar distribution of transporters. The use of in silico models to analyze the in vitro data related to hepatotoxic activity of acetaminophen (APAP) demonstrated the advantage of the integration of kinetic and dynamic aspects for a better understanding of the in vitro cell behavior. The bioactivation of APAP and its related cytotoxicity was assessed in a system compatible to high-throughput screening. The approach also proved to be a good strategy to reduce the time necessary to obtain fully differentiated cell cultures. In conclusion, HepaRG cells cultured in 3D spinner-bioreactors are an attractive tool for toxicological studies, showing a liver-like performance and demonstrating a practical applicability for toxicodynamic approaches.
authors: Instituto de Biologia Experimental e Tecnológica, 2781-901 Oeiras, Portugal.
authors: Sofia B
authors: SB
authors: Leite
authors: Iwona
authors: I
authors: Wilk-Zasadna
authors: Jose M
authors: JM
authors: Zaldivar
authors: Elodie
authors: E
authors: Airola
authors: Marcos A
authors: MA
authors: Reis-Fernandes
authors: Milena
authors: M
authors: Mennecozzi
authors: Christiane
authors: C
authors: Guguen-Guillouzo
authors: Christopher
authors: C
authors: Chesne
authors: Claude
authors: C
authors: Guillou
authors: Paula M
authors: PM
authors: Alves
authors: Sandra
authors: S
authors: Coecke
mesh_terms: D000082:Acetaminophen / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D000418:Albumins / Q000378:metabolism; D018712:Analgesics, Non-Narcotic / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D000826:Animal Testing Alternatives / Q000379:methods*; D019149:Bioreactors; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D003198:Computer Simulation; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis; D004790:Enzyme Induction / Q000187:drug effects; D014453:Glucuronosyltransferase / Q000378:metabolism; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D018675:Toxicity Tests
chemical_list: D000418:Albumins; D018712:Analgesics, Non-Narcotic; D000082:Acetaminophen; D051544:Cytochrome P-450 CYP3A; D014453:Glucuronosyltransferase