Cold preservation of the liver with oxygenation by a two-layer method.
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.
Odaira
Masanori
M
Division of General and Gastroenterological Surgery, Department of Surgery, School of Medicine, Showa University, Tokyo, Japan. ij100106-8463@tbk.t-com.ne.jp
Aoki
Takeshi
T
Miyamoto
Yoichi
Y
Yasuhara
Rika
R
Jin
Zhenghao
Z
Yu
Jiwei
J
Nishino
Nobukazu
N
Yamada
Kousuke
K
Kusano
Tomokazu
T
Hayashi
Ken
K
Yasuda
Daisuke
D
Koizumi
Tomotake
T
Mitamura
Keitaro
K
Enami
Yuta
Y
Niiya
Takashi
T
Murai
Noriyuki
N
Kato
Hirohisa
H
Shimizu
Yoshinori
Y
Kamijyo
Ryutaro
R
Kusano
Mitsuo
M
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
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