Hepatobiliary disposition in primary cultures of dog and monkey hepatocytes.
Hepatobiliary transporters are a major route for elimination of xenobiotics and endogenous products. In vitro hepatobiliary models have been reported for human and rat, but not for the other preclinical species used in safety evaluation. We have established methodologies for culturing dog and monkey hepatocytes with optimal bile canalicular formation and function, using a sandwich culture comprising rigid collagen substratum and gelled collagen overlay. Hepatic uptake utilizing sinusoidal transporters and biliary excretion through canalicular transporters were assessed using the bile salt taurocholate, salicylate (negative control), and the Bsep inhibitors cyclosporin A (CsA) and glyburide. There was significant taurocholate and salicylate canalicular efflux in dog and monkey hepatocytes, although the amount of salicylate transported was one thousandth that of taurocholate. Species differences were observed, as glyburide significantly inhibited taurocholate uptake in monkey (64% at 10 microM) but not dog hepatocytes, and inhibited taurocholate efflux in dog (100% at 10 microM) but not monkey hepatocytes. CsA did not inhibit bile salt uptake and significantly inhibited canalicular efflux in dog (at 0.1 microM) and monkey (at 1 and 10 microM) hepatocyte cultures. These results suggest that glyburide is a bile salt uptake inhibitor in monkey but not in dog hepatocytes and that CsA inhibits bile salt canalicular efflux but not basolateral uptake in these species. We have established dog and monkey hepatocytes in sandwich culture with intact bile canalicular formation and function. The differences observed in taurocholate transport between dog and monkey hepatocytes may be indicative of in vivo species differences.
Rose
Kelly A
KA
Department of Pharmacokinetics, Pharmacodynamics, and Metabolism and Drug Safety Evaluation, Pfizer Global Research and Development, Ann Arbor, Michigan 48105, USA.
Kostrubsky
Vsevolod
V
Sahi
Jasminder
J
D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism*; D001692:Biological Transport; D018929:Cell Culture Techniques / Q000379:methods; D003094:Collagen / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D004285:Dogs; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D005905:Glyburide / Q000494:pharmacology; D000882:Haplorhini; D022781:Hepatocytes / Q000378:metabolism*; D008658:Inactivation, Metabolic; D020156:Salicylic Acid / Q000493:pharmacokinetics; D013045:Species Specificity; D013656:Taurocholic Acid / Q000493:pharmacokinetics
D001647:Bile Acids and Salts; D013656:Taurocholic Acid; D016572:Cyclosporine; D003094:Collagen; D020156:Salicylic Acid; D005905:Glyburide