Use of cassette dosing in sandwich-cultured rat and human hepatocytes to identify drugs that inhibit bile acid transport

Toxicol In Vitro. 2010 Feb;24(1):297-309. doi: 10.1016/j.tiv.2009.08.009. Epub 2009 Aug 23.


Hepatocellular accumulation of bile acids due to inhibition of the canalicular bile salt export pump (BSEP/ABCB11) is one proposed mechanism of drug-induced liver injury (DILI). Some hepatotoxic compounds also are potent inhibitors of bile acid uptake by Na(+)-dependent taurocholate cotransporting polypeptide (NTCP/SLC10A1). This study used a cassette dosing approach in rat and human sandwich-cultured hepatocytes (SCH) to determine whether known or suspected hepatotoxic drugs inhibit bile acid transport individually or in combination. [(3)H]-Taurocholate served as the NTCP/BSEP probe substrate. Individually, cyclosporin A and rifampin decreased taurocholate in vitro biliary clearance (Cl(biliary)) and biliary excretion index (BEI) by more than 20% in rat SCH, suggesting that these drugs primarily inhibited canalicular efflux. In contrast, ampicillin, carbenicillin, cloxacillin, nafcillin, oxacillin, carbamazepine, pioglitazone, and troglitazone decreased the in vitro Cl(biliary) by more than 20% with no notable change in BEI, suggesting that these drugs primarily inhibited taurocholate uptake. Cassette dosing (n=2-4 compounds per cassette) in rat SCH yielded similar findings, and results in human SCH were consistent with rat SCH. In summary, cassette dosing in SCH is a useful in vitro approach to identify compounds that inhibit the hepatic uptake and/or excretion of bile acids, which may cause DILI.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Infective Agents / pharmacology
  • Bile / metabolism
  • Bile Acids and Salts / metabolism*
  • Biological Transport, Active / drug effects
  • Cells, Cultured
  • Cholagogues and Choleretics / pharmacology*
  • Cyclosporine / pharmacology
  • Data Interpretation, Statistical
  • Dose-Response Relationship, Drug
  • Drug Evaluation, Preclinical / methods
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism*
  • Humans
  • Hypoglycemic Agents / pharmacology
  • Immunosuppressive Agents / pharmacology
  • Liver / metabolism
  • Male
  • Rats
  • Rats, Wistar
  • Taurocholic Acid / metabolism


  • Anti-Infective Agents
  • Bile Acids and Salts
  • Cholagogues and Choleretics
  • Hypoglycemic Agents
  • Immunosuppressive Agents
  • Taurocholic Acid
  • Cyclosporine