title: The utility of cold-preserved human hepatocytes in studies on cytochrome P450 induction and hepatic drug transport.
abstract: Human hepatocytes that had been cold-preserved in SureTran(TM) matrix (Abcellute Ltd, Cardiff, UK) were used for studies on cell viability, cytochrome P450 (CYP) 3A4, 2B6 and 1A2 induction and hepatic drug transporters. It has recently been shown that basal CYP activities are maintained in cold-preserved hepatocytes (Palmgren et al., 2012). After 5 d of cold preservation, the viability was still more than 70%, and after 8 d it was around 60%. In hepatocytes that had been cold-preserved for 3 d, the activity of CYP3A4 was induced around 15-fold upon treatment with 8 µM rifampicin for 72 h. For CYP2B6, the activity was induced 4- to 16-fold in hepatocytes that had been cold-preserved for 3 d and thereafter treated with 1 mM phenobarbital for 72 h. The activity of CYP1A2 was low and close to the limit of detection in non-treated cells that had been cold-preserved for up to 3 d, while the activity increased in cells treated with 0.3-25 µM β-naphthoflavone for 72 h. CYP3A4, 2B6 and 1A2 mRNA levels were only determined with hepatocytes from one donor and increased upon treatment with the inducers. Hepatic uptakes of estrone-3-sulfate, taurocholate, ipratropium and rosuvastatin were stable in human hepatocytes that had been cold-preserved for up to 2 d. In summary, cold-preserved human hepatocytes demonstrate retained viability and can advantageously be used for in vitro induction studies and for studies of hepatic uptake transporters.
authors.affiliation: DMPK, CNSP iMed, AstraZeneca R&D Södertälje, Södertälje, Sweden.
authors.forename: Sanja
authors.initials: S
authors.lastname: Juric
authors.forename: Patrik
authors.initials: P
authors.lastname: Lundquist
authors.forename: Yin
authors.initials: Y
authors.lastname: Hu
authors.forename: Anna
authors.initials: A
authors.lastname: Juréus
authors.forename: Anna-Karin
authors.initials: AK
authors.lastname: Sohlenius-Sternbeck
mesh_terms: D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism*; D004970:Estrone / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D005464:Fluorobenzenes / Q000493:pharmacokinetics; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D009241:Ipratropium / Q000493:pharmacokinetics; D008099:Liver / Q000378:metabolism*; D027361:Organic Anion Transporters / Q000378:metabolism / Q000502:physiology; D010634:Phenobarbital / Q000494:pharmacology; D011743:Pyrimidines / Q000493:pharmacokinetics; D012333:RNA, Messenger / Q000378:metabolism; D012293:Rifampin / Q000493:pharmacokinetics / Q000494:pharmacology; D000068718:Rosuvastatin Calcium; D013449:Sulfonamides / Q000493:pharmacokinetics; D013656:Taurocholic Acid / Q000493:pharmacokinetics
chemical_list: D005464:Fluorobenzenes; D027361:Organic Anion Transporters; D011743:Pyrimidines; D012333:RNA, Messenger; D013449:Sulfonamides; D004970:Estrone; D013656:Taurocholic Acid; D000068718:Rosuvastatin Calcium; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D009241:Ipratropium; C017296:estrone sulfate; D012293:Rifampin; D010634:Phenobarbital