title: Induction, expression and maintenance of cytochrome P450 isoforms in long-term cultures of primary human hepatocytes.
abstract: Within the past decade, tremendous progress has been made in the isolation and culture of human hepatocytes for drug metabolism and toxicology, which could potentially reduce the number of animal experiments performed. However, human hepatocyte cultures are still not widely used for preclinical drug testing, partly due to inconsistent supply and quality of human tissue. Thus, the aim of this study was to evaluate primary cultured human hepatocytes from different patients over a study period of 14 days, by assays that characterise cell quality and function. We found urea production and albumin synthesis in all cell cultures over at least 7 days. Cytochrome P4501A2, CYP2D6, and CYP3A4 protein expression was demonstrated by Western Blot analysis and CYP1A1/2 and CYP3A4 induction by 3-methylcholantrene, phenobarbital or rifampicin over 14 days. In addition, we saw that UDP-glucoronyltransferase activity was preserved in human hepatocytes over 2 weeks. In conclusion, we could show that primary human hepatocytes isolated from discarded liver tissue can consistently be kept in culture over a long time period and are therefore well suited for preclinical drug testing.
authors.affiliation: Department General, Visceral and Transplantation Surgery, Charité Campus Virchow-Klinikum, Universitätsmedizin Berlin, Humboldt University of D-Berlin, Germany.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
authors.forename: Liegang
authors.initials: L
authors.lastname: Liu
authors.forename: Masashi
authors.initials: M
authors.lastname: Shimada
authors.forename: Aiguo
authors.initials: A
authors.lastname: Wang
authors.forename: Maren
authors.initials: M
authors.lastname: Ruhnke
authors.forename: Peter
authors.initials: P
authors.lastname: Heeckt
authors.forename: Andrea R
authors.initials: AR
authors.lastname: Muller
authors.forename: Natascha C
authors.initials: NC
authors.lastname: Nussler
authors.forename: Peter
authors.initials: P
authors.lastname: Neuhaus
authors.forename: Andreas
authors.initials: A
authors.lastname: Nussler
mesh_terms: D000418:Albumins / Q000378:metabolism; D000826:Animal Testing Alternatives; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004790:Enzyme Induction / Q000187:drug effects; D005947:Glucose / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D014116:Toxicology / Q000379:methods*; D014508:Urea / Q000378:metabolism
chemical_list: D000418:Albumins; D007527:Isoenzymes; D019344:Lactic Acid; D013739:Testosterone; D014508:Urea; D003577:Cytochrome P-450 Enzyme System; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; D005947:Glucose