title: Three-dimensional co-culture of primary human liver cells in bioreactors for in vitro drug studies: effects of the initial cell quality on the long-term maintenance of hepatocyte-specific functions.
abstract: In vitro culture models that employ human liver cells could be potent tools for predictive studies on drug toxicity and metabolism in the pharmaceutical industry. A bioreactor culture model was developed that permits the three-dimensional co-culture of liver cells under continuous medium perfusion with decentralised mass exchange and integral oxygenation. We tested the ability of the system to support the long-term maintenance and differentiation of primary human liver cells. The effects of the initial cell quality were investigated by comparing cultures from resected, non-preserved liver with cultures from liver graft tissue damaged by long-term preservation. In cultures originating from non-preserved liver, protein and urea synthesis, glucose metabolism, and cytochrome (CYP450) activities were stable over the 2-week culture period, with maximal activities at the end of the first week in culture. Enzyme induction led to increased 7-ethoxyresorufin O-deethylase activities of up to 20 times the basal value. In cultures from preservation-damaged liver, recovery of metabolic activities was detected during bioreactor culture. After two weeks, most biochemical parameters approached those of cultures from non-preserved human liver. Light microscopy demonstrated the three-dimensional reorganisation of hepatocytes and non-parenchymal cells in co-culture. Long-term maintenance, and even the regeneration of specific functional activities of human liver cells, can be achieved in the bioreactor. This could facilitate the introduction into the pharmaceutical industry of in vitro drug testing with primary human liver cells.
authors.affiliation: Department of Experimental Surgery, Surgical Clinic, Charité Campus Virchow-Klinikum, Humboldt University of Berlin, Augustenburger Platz 1, 13353 Berlin, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Zeilinger
authors.forename: Igor M
authors.initials: IM
authors.lastname: Sauer
authors.forename: Gesine
authors.initials: G
authors.lastname: Pless
authors.forename: Catrin
authors.initials: C
authors.lastname: Strobel
authors.forename: Jeannette
authors.initials: J
authors.lastname: Rudzitis
authors.forename: Aiguo
authors.initials: A
authors.lastname: Wang
authors.forename: Andres K
authors.initials: AK
authors.lastname: Nüssler
authors.forename: Alexander
authors.initials: A
authors.lastname: Grebe
authors.forename: Lei
authors.initials: L
authors.lastname: Mao
authors.forename: Stefan H G
authors.initials: SH
authors.lastname: Auth
authors.forename: Juliane
authors.initials: J
authors.lastname: Unger
authors.forename: Peter
authors.initials: P
authors.lastname: Neuhaus
authors.forename: Jörg C
authors.initials: JC
authors.lastname: Gerlach
mesh_terms: D000418:Albumins / Q000378:metabolism; D000826:Animal Testing Alternatives / Q000295:instrumentation / Q000379:methods*; D001219:Aspartate Aminotransferases / Q000378:metabolism; D019149:Bioreactors; D002245:Carbon Dioxide / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D018920:Coculture Techniques; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008012:Lidocaine / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D016031:Liver Transplantation; D009926:Organ Preservation; D010100:Oxygen / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D014508:Urea / Q000378:metabolism
chemical_list: D000418:Albumins; D002245:Carbon Dioxide; D013739:Testosterone; D014508:Urea; D008012:Lidocaine; D007770:L-Lactate Dehydrogenase; D019363:Cytochrome P-450 CYP1A1; D001219:Aspartate Aminotransferases; D005947:Glucose; D010100:Oxygen