title: Cold preservation of the liver with oxygenation by a two-layer method.
abstract: BACKGROUND
  The two-layer method (TLM) has recently been found to be superior to simple cold storage in University of Wisconsin (UW) solution as a means of pancreas preservation for islet transplantation. In this study, we investigated whether TLM would result in better hepatocyte function over UW cold storage and if it could be applied to hepatocyte transplantation.
  
  
  METHODS
  Hepatocytes from male Sprague Dawley rat livers were isolated and divided into three groups: a non-preservation group (group 1), a 10-h preservation group (group 2), and a 24-h preservation group (group 3). Groups 2 and 3 were then divided into three subgroups: a group preserved by the TLM (subgroup a), a group preserved in UW solution (subgroup b), and a group preserved in water (subgroup c). Isolated hepatocytes were evaluated for cell yield, viability, and adenosine triphosphate level after preservation. Hepatocytes were either cultured or transplanted.
  
  
  RESULTS
  Although no differences in cell yield or morphological findings were observed between any of the groups, TLM significantly improved hepatocyte viability and adenosine triphosphate levels in comparison with UW cold storage. Albumin production or urea synthesis were significantly higher in subgroup 3a than in subgroup 3b at almost all time points. Surprisingly, after hepatocyte transplantation, the serum albumin level in subgroup 2a was significantly higher than in subgroup 2b at every time point.
  
  
  CONCLUSIONS
  The results of this study demonstrated that liver preservation by the TLM before hepatocyte isolation might be beneficial and will be useful in the field of hepatotocyte transplantation.
authors.affiliation: Division of General and Gastroenterological Surgery, Department of Surgery, School of Medicine, Showa University, Tokyo, Japan. ij100106-8463@tbk.t-com.ne.jp
authors.forename: Masanori
authors.initials: M
authors.lastname: Odaira
authors.forename: Takeshi
authors.initials: T
authors.lastname: Aoki
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Miyamoto
authors.forename: Rika
authors.initials: R
authors.lastname: Yasuhara
authors.forename: Zhenghao
authors.initials: Z
authors.lastname: Jin
authors.forename: Jiwei
authors.initials: J
authors.lastname: Yu
authors.forename: Nobukazu
authors.initials: N
authors.lastname: Nishino
authors.forename: Kousuke
authors.initials: K
authors.lastname: Yamada
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Kusano
authors.forename: Ken
authors.initials: K
authors.lastname: Hayashi
authors.forename: Daisuke
authors.initials: D
authors.lastname: Yasuda
authors.forename: Tomotake
authors.initials: T
authors.lastname: Koizumi
authors.forename: Keitaro
authors.initials: K
authors.lastname: Mitamura
authors.forename: Yuta
authors.initials: Y
authors.lastname: Enami
authors.forename: Takashi
authors.initials: T
authors.lastname: Niiya
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Murai
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Kato
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Shimizu
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Kamijyo
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Kusano
mesh_terms: D000241:Adenosine; D000255:Adenosine Triphosphate / Q000378:metabolism; D000493:Allopurinol; D000818:Animals; D018929:Cell Culture Techniques; D003080:Cold Temperature; D005978:Glutathione; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D051537:Hepatocyte Nuclear Factor 1 / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology* / Q000637:transplantation; D007328:Insulin; D008099:Liver / Q000166:cytology / Q000502:physiology*; D016031:Liver Transplantation / Q000379:methods*; D008297:Male; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D011887:Raffinose; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012709:Serum Albumin / Q000096:biosynthesis / Q000235:genetics
chemical_list: D007328:Insulin; D019188:Organ Preservation Solutions; D012709:Serum Albumin; C055206:University of Wisconsin-lactobionate solution; D051537:Hepatocyte Nuclear Factor 1; D012313:RNA; D000493:Allopurinol; D000255:Adenosine Triphosphate; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose