Co-culture of Hepatocytes and Kupffer Cells as an In Vitro Model of Inflammation and Drug-Induced Hepatotoxicity

• Published in: Journal of pharmaceutical sciences Vol. 105; no. 2; pp. 950 - 964
• Main Authors: Rose, Kelly A., Holman, Natalie S., Green, Angela M., Andersen, Melvin E., LeCluyse, Edward L.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.02.2016
• Subjects: Animals; Cells, Cultured; Chemical and Drug Induced Liver Injury - metabolism; co-culture; Coculture Techniques; Dose-Response Relationship, Drug; Glucocorticoids - toxicity; hepatocytes; Hepatocytes - drug effects; Hepatocytes - metabolism; hepatotoxicity; Inflammation - chemically induced; Inflammation - metabolism; Inflammation Mediators - metabolism; in vitro model systems; Kupffer cells; Kupffer Cells - drug effects; Kupffer Cells - metabolism; Lipopolysaccharides - toxicity; Male; Rats; Rats, Sprague-Dawley; co-culture; hepatotoxicity; hepatocytes; Kupffer cells; in vitro model systems
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1016/S0022-3549(15)00192-6

Immune-mediated drug-induced hepatotoxicity is often unrecognized as a potential mode of action due to the lack of appropriate in vitro models. We have established an in vitro rat donor-matched hepatocyte and Kupffer cell co-culture (HKCC) model to study immune-related responses to drug exposure. Optimal cell culture conditions were identified for the maintenance of co-cultures based on cell longevity, monolayer integrity, and cytokine response after lipopolysaccharide (LPS) exposure. Hepatocyte monocultures and HKCCs were then used to test a subset of compounds associated with hepatotoxic effects with or without LPS. Cytokine levels and metabolic activity (cytochrome P450 3A [Cyp3A]) were measured after a 48-h exposure to monitor endotoxin-induced changes in acute phase and functional end points. LPS-activated HKCCs, but not hepatocyte monocultures, treated with trovafloxacin or acetaminophen, compounds associated with immune-mediated hepatotoxicity, showed LPS-dependent decreases in interleukin-6 production with concomitant increases in Cyp3A activity. Differential endotoxin- and model-dependent alterations were observed in cytokine profiles and Cyp3A activity levels that corresponded to specific compounds. These results indicate the utility of the HKCC model system to discern compound-specific effects that may lead to enhanced or mitigate hepatocellular injury due to innate or adaptive immune responses.