title: Comparison of culture media for bile Acid transport studies in primary human hepatocytes.
abstract: BACKGROUND
  Primary human hepatocytes are a useful in vitro model system to examine hepatic biochemical pathways, liver disorders and/or pharmacotherapies. This system can also be used for transport studies to investigate uptake and excretion of bile acids. Proper modeling of hepatic function requires careful attention to media components, and culture substrates and conditions.
  
  
  OBJECTIVE
  To examine the effects of different culture media and conditions on bile acid transport in cultured human hepatocytes.
  
  
  RESULTS
  Hepatocytes cultured in Williams' medium E showed an increase in both uptake and excretion of taurocholate compared to cells cultured in Dulbecco's Modified Eagle Medium (DMEM). Supplementation of DMEM with glutathione or ascorbic acid did not compensate for the lower transport. The difference can be explained by lower mRNA expression of the transporter proteins sodium taurocholate cotransporting polypeptide (NTCP) and bile salt export pump (BSEP; ABCB11) when cultured in DMEM. Hepatocytes cultured in DMEM also display fewer and smaller bile canaliculi. Following extended time in culture supplementation of Williams' medium E with dexamethasone increased the expression of NTCP and BSEP.
  
  
  CONCLUSIONS
  Williams' medium E is superior to DMEM for transport studies in primary human hepatocytes. Supplementation with dexamethasone increase mRNA levels of NTCP and BSEP.
authors.affiliation: Division of Pediatrics, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm 14186, Sweden ; Division of Transplantation Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm 14186, Sweden ; Department of Laboratory Medicine, Karolinska Institutet, Stockholm 14186, Sweden.
authors.forename: Lisa-Mari
authors.initials: LM
authors.lastname: Mörk
authors.affiliation: Division of Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm 14186, Sweden.
authors.forename: Bengt
authors.initials: B
authors.lastname: Isaksson
authors.affiliation: Division of Transplantation Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm 14186, Sweden.
authors.forename: Nicola
authors.initials: N
authors.lastname: Boran
authors.affiliation: Division of Transplantation Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm 14186, Sweden.
authors.forename: Bo-Göran
authors.initials: BG
authors.lastname: Ericzon
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Stephen
authors.initials: S
authors.lastname: Strom
authors.affiliation: Division of Pediatrics, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm 14186, Sweden.
authors.forename: Björn
authors.initials: B
authors.lastname: Fischler
authors.affiliation: Division of Transplantation Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm 14186, Sweden.
authors.forename: Ewa
authors.initials: E
authors.lastname: Ellis
keywords: AA, ascorbic acid; BSEP; BSEP, bile salt export pump; CgamF, cholylglycylamido-fluorescein; DMEM, Dulbecco's Modified Eagle Medium; GSH, glutathione; HBSS, Hank's Balanced Salt Solution; MRP2, multidrug resistance protein 2; NTCP; NTCP, sodium taurocholate cotransporting polypeptide; OATP, organic anion-transporting polypeptide; WE, Williams' medium E; bile acid transport; dexamethasone; primary human hepatocytes