title: Evolution of graft morphology and function after recellularization of decellularized rat livers.
abstract: Decellularization of livers is a well-established procedure. Data on different reseeding techniques or the functional evolution and reorganization processes of repopulated grafts remains limited. A proprietary, customized bioreactor was established to repopulate decellularized rat livers (n = 21) with primary rat hepatocytes (150 × 106cells) via the hepatic artery and to subsequently evaluate graft morphology and function during 7 days of ex vivo perfusion. Grafts were analysed at 1 h, 6 h, 12 h, 24 h, 3 days, 5 days and 7 days after recellularization (all n = 3) by immunohistological evaluation, hepatocyte-related enzyme (aspartate transaminase, alanine transaminase and lactate dehydrogenase) and albumin measurement in the perfusate. This appears to be the first available protocol for repopulation of rat livers via the hepatic artery. Within the first 24 h after repopulation, the hepatocytes seemed to migrate out of the vascular network and form clusters in the parenchymal space around the vessels. Graft function increased for the first 24 h after repopulation with a significantly higher function compared to standard two-dimensional culture after 24 h. Thereafter, graft function constantly decreased with significantly lower values after 6 days and 7 days of perfusion, although histologically viable hepatocytes were found even after this period. The data suggests that, owing to a constant loss of function, repopulated grafts should potentially be implanted as soon as cell engraftment and graft re-organization are completed. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Butter
authors.affiliation: Department of General, Visceral and Transplantation Surgery, Johannes Gutenberg Universitaet, Mainz, Germany.
authors.forename: Khalid
authors.initials: K
authors.lastname: Aliyev
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Karl-Herbert
authors.initials: KH
authors.lastname: Hillebrandt
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Nathanael
authors.initials: N
authors.lastname: Raschzok
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Kluge
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Nicolai
authors.initials: N
authors.lastname: Seiffert
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Tang
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Napierala
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Ashraf I
authors.initials: AI
authors.lastname: Muhamma
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Reutzel-Selke
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Andreou
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Johann
authors.initials: J
authors.lastname: Pratschke
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Igor M
authors.initials: IM
authors.lastname: Sauer
authors.affiliation: Department of Surgery, Campus Virchow-Klinikum and Campus Charité Mitte, Charité Universitaetsmedizin Berlin, Berlin, Germany.|Berlin Institute of Health (BIH), Berlin, Germany.
authors.forename: Benjamin
authors.identifier: 0000-0002-4205-9890
authors.initials: B
authors.lastname: Struecker
mesh_terms: D000410:Alanine Transaminase / Q000378:metabolism; D000418:Albumins / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D019149:Bioreactors; D002470:Cell Survival; D005260:Female; D005947:Glucose / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D011917:Rats, Inbred Lew; D019737:Transplants / Q000502:physiology*
chemical_list: D000418:Albumins; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D005947:Glucose; D011188:Potassium
keywords: arterial perfusion; bioreactor; ex vivo perfusion; graft analysis; liver decellularization; liver recellularization