title: Oxygenated Static Preservation of Donation after Cardiac Death Liver Grafts Improves Hepatocyte Viability and Function.
abstract: BACKGROUND
  Cell therapy, such as hepatocyte transplantation (HTx), is promising for the treatment of metabolic liver diseases or as a bridge to orthotopic liver transplantation in patients with fulminant liver failure. However, one of the limitations of this therapy is the shortage of donors. The present study aims to investigate whether the two-layer method (TLM) of cold preservation with oxygenation improves the viability and activity of hepatocytes from rat donation after cardiac death (DCD) donors compared with results obtained with the University of Wisconsin (UW) solution. Moreover, we evaluated the hepatocyte function after culture or transplantation into the spleen.
  
  
  METHODS
  We used male Sprague-Dawley rats for this study. The DCD model was induced by phrenotomy after injecting heparin. We assigned rats based on warm ischemia times of 15 and 30 min to groups S and L, respectively. Each group (n = 5) was then subdivided as follows: (1) group S: not preserved (S/N), preserved by TLM for 3 h (S/TLM3) and 12 h (S/TLM12), and in the UW solution for 3 h (S/UW3) and 12 h (S/UW12), and (2) group L: not preserved (L/N), preserved by TLM for 3 h (L/TLM3) and 12 h (L/TLM12), and in the UW solution for 3 h (L/UW3) and 12 h (L/UW12). The cell viability and function of isolated DCD hepatocytes were analyzed for culture or HTx into the spleen.
  
  
  RESULTS
  The viability and ATP levels of DCD hepatocytes significantly improved after TLM compared with the values after preservation in cold UW solution in group S/N (p < 0.059). The levels of albumin production and urea synthesis by hepatocytes after culture were significantly higher in groups S/TLM3 and S/TLM12 than in groups S/UW3 and S/UW12 (p < 0.05), respectively. Further, serum albumin levels after HTx were also markedly higher in groups S/TLM3 and S/TLM12 than in groups S/UW3 and S/UW12. The morphological features revealed that cultured and transplanted hepatocytes remained clearly viable and maintained an expression for specific hepatic function, such as the production of albumin and glycogen.
  
  
  CONCLUSIONS
  This novel method of oxygenated cold preservation of DCD livers can expand the hepatocyte donor pool for HTx and establish a wider application of this developing technique.
authors.affiliation: Division of General and Gastroenterological Surgery, Department of Surgery, School of Medicine, Showa University, Tokyo, Japan.
authors.forename: Jiwei
authors.initials: J
authors.lastname: Yu
authors.forename: Masahiko
authors.initials: M
authors.lastname: Murakami
authors.forename: Takeshi
authors.initials: T
authors.lastname: Aoki
authors.forename: Bojian
authors.initials: B
authors.lastname: Jiang
authors.forename: Zhenghao
authors.initials: Z
authors.lastname: Jin
authors.forename: Tomotake
authors.initials: T
authors.lastname: Koizumi
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Kusano
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Kamijo
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Miyamoto
authors.forename: Yuta
authors.initials: Y
authors.lastname: Enami
authors.forename: Makoto
authors.initials: M
authors.lastname: Watanabe
authors.forename: Koji
authors.initials: K
authors.lastname: Otsuka
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D003080:Cold Temperature; D003643:Death; D022781:Hepatocytes / Q000502:physiology*; D016031:Liver Transplantation; D008297:Male; D009926:Organ Preservation / Q000379:methods*; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000418:Albumins; D000255:Adenosine Triphosphate; D010100:Oxygen