title: Bioactivation and toxicity of acetaminophen in a rat hepatocyte micropatterned coculture system.
abstract: We have recently shown that primary rat hepatocytes organized in micropatterned cocultures with murine embryonic fibroblasts (HepatoPac™) maintain high levels of liver functions for at least 4 weeks. In this study, rat HepatoPac was assessed for its utility to study chemical bioactivation and associated hepatocellular toxicity. Treatment of HepatoPac cultures with acetaminophen (APAP) over a range of concentrations (0-15 mM) was initiated at 1, 2, 3, or 4 weeks followed by the assessment of morphological and functional endpoints. Consistent and reproducible concentration-dependent effects on hepatocyte structure, viability, and basic functions were observed over the 4-week period, and were exacerbated by depleting glutathione using buthionine sulfoximine or inducing CYP3A using dexamethasone, presumably due to increased reactive metabolite-induced stress and adduct formation. In conclusion, the results from this study demonstrate that rat HepatoPac represents a structurally and functionally stable hepatic model system to assess the long-term effects of bioactivated compounds.
authors.affiliation: Hepregen Corporation, Medford, MA, 02155, U.S.A.
authors.forename: Okechukwu
authors.initials: O
authors.lastname: Ukairo
authors.forename: Michael
authors.initials: M
authors.lastname: McVay
authors.forename: Stacy
authors.initials: S
authors.lastname: Krzyzewski
authors.forename: Simon
authors.initials: S
authors.lastname: Aoyama
authors.forename: Kelly
authors.initials: K
authors.lastname: Rose
authors.forename: Melvin E
authors.initials: ME
authors.lastname: Andersen
authors.forename: Salman R
authors.initials: SR
authors.lastname: Khetani
authors.forename: Edward L
authors.initials: EL
authors.lastname: Lecluyse
mesh_terms: D000082:Acetaminophen / Q000633:toxicity*; D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D000418:Albumins / Q000378:metabolism; D018712:Analgesics, Non-Narcotic / Q000633:toxicity*; D000818:Animals; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D018920:Coculture Techniques; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology; D005978:Glutathione / Q000037:antagonists & inhibitors* / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D061251:Primary Cell Culture; D051381:Rats; D017207:Rats, Sprague-Dawley; D015854:Up-Regulation / Q000187:drug effects; D014508:Urea / Q000378:metabolism
chemical_list: D000418:Albumins; D018712:Analgesics, Non-Narcotic; D000082:Acetaminophen; D019328:Buthionine Sulfoximine; D003907:Dexamethasone; D000255:Adenosine Triphosphate; D014508:Urea; D051544:Cytochrome P-450 CYP3A; D005978:Glutathione
keywords: Acetaminophen; Bioactivation; Cocultures; Micropatterning; Rat Hepatocytes