title: Explanted diseased livers - a possible source of metabolic competent primary human hepatocytes.
abstract: Being an integral part of basic, translational and clinical research, the demand for primary human hepatocytes (PHH) is continuously growing while the availability of tissue resection material for the isolation of metabolically competent PHH remains limited. To overcome current shortcomings, this study evaluated the use of explanted diseased organs from liver transplantation patients as a potential source of PHH. Therefore, PHH were isolated from resected surgical specimens (Rx-group; n = 60) and explanted diseased livers obtained from graft recipients with low labMELD-score (Ex-group; n = 5). Using established protocols PHH were subsequently cultured for a period of 7 days. The viability and metabolic competence of cultured PHH was assessed by the following parameters: morphology and cell count (CyQuant assay), albumin synthesis, urea production, AST-leakage, and phase I and II metabolism. Both groups were compared in terms of cell yield and metabolic function, and results were correlated with clinical parameters of tissue donors. Notably, cellular yields and viabilities were comparable between the Rx- and Ex-group and were 5.3±0.5 and 2.9±0.7×106 cells/g liver tissue with 84.3±1.3 and 76.0±8.6% viability, respectively. Moreover, PHH isolated from the Rx- or Ex-group did not differ in regards to loss of cell number in culture, albumin synthesis, urea production, AST-leakage, and phase I and II metabolism (measured by the 7-ethoxycoumarin-O-deethylase and uracil-5'-diphosphate-glucuronyltransferase activity). Likewise, basal transcript expressions of the CYP monooxygenases 1A1, 2C8 and 3A4 were comparable as was their induction when treated with a cocktail that consisted of 3-methylcholantren, rifampicin and phenobarbital, with increased expression of CYP 1A1 and 3A4 mRNA while transcript expression of CYP 2C8 was only marginally changed. In conclusion, the use of explanted diseased livers obtained from recipients with low labMELD-score might represent a valuable source of metabolically competent PHH which are comparable in viability and function to cells obtained from specimens following partial liver resection.
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Kleine
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Riemer
authors.affiliation: Institute of Pathology, Hannover Medical School, Hannover, Germany.
authors.forename: Till
authors.initials: T
authors.lastname: Krech
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Daphne
authors.initials: D
authors.lastname: DeTemple
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Jäger
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Lehner
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany; German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Hannover, Germany.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Manns
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Klempnauer
authors.affiliation: Center of Pharmacology and Toxicology, Hannover Medical School, Hannover, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Borlak
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Hueseyin
authors.initials: H
authors.lastname: Bektas
authors.affiliation: ReMediES, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany; German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Hannover, Germany.
authors.forename: Florian W R
authors.initials: FW
authors.lastname: Vondran
mesh_terms: D000328:Adult; D000368:Aged; D000418:Albumins / Q000096:biosynthesis / Q000235:genetics; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics; D002452:Cell Count; D002470:Cell Survival; D005260:Female; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008099:Liver / Q000473:pathology* / Q000601:surgery; D008107:Liver Diseases / Q000473:pathology*; D016031:Liver Transplantation; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014508:Urea / Q000378:metabolism; D055815:Young Adult
chemical_list: D000418:Albumins; D012333:RNA, Messenger; D014508:Urea; D001189:Aryl Hydrocarbon Hydroxylases