Grant-in-Aid for General Scientific Research (A)
|Allocation Type||Single-year Grants|
|Research Institution||Hokkaido University|
UCHINO Junichi Hokkaido University School of Medicine, Professor, 医学部, 教授 (40000989)
MATUSHITA Michiaki Hokkaido University School of Medicine, Lecturer, 医学部・附属病院, 講師 (20250425)
NAKAJIMA Yasuaki Hokkaido University School of Medicine, Lecturer, 医学部・附属病院, 講師 (90198101)
KUZUMAKI Noboru Hokkaido University School of Medicine, Professor, 医学部, 教授 (80091445)
|Project Period (FY)
1991 – 1993
Completed(Fiscal Year 1993)
|Budget Amount *help
¥21,800,000 (Direct Cost : ¥21,800,000)
Fiscal Year 1993 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 1992 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 1991 : ¥19,800,000 (Direct Cost : ¥19,800,000)
|Keywords||Hybrid artificial liver system / Culture of hepatocyte / Hepatic failure / Natural antibody / Human hepatocyte / Warm ischemia / Hepatic function / ヒト肝細胞培養 / 酸素分圧 / バイオリアクター / 培養ブタ肝細胞 / ブタ肝細胞培養 / アミノ酸代謝|
For the clinical application of our hybrid artificial liver system, multiplated hepatocyte monolayrs, the immunological examination on au use of xenogeneic hepatocytes, the methods of isolation and culture of human gepatocytes, and efficacy of the artificial liver system on anhepatic xenogeneic animals were studied.
1. The basic studies on culture of xenogeneic hepatocytes
(1)The functions of cultured porcine hepatocytes had functioned well for 5 days in the plasma from hepatic failure patients. The high levels of the plasma phenylalanine and methonine were reduced significangtly and the molar ratio was improved by the cultured hepatocytes.
(2)Among 103 human sera, seven (6.8%) sera showed cytaotoxicity to cultured porcine hepatocytes. These cytotoxic reactions were thought to be mediated by natural antibodies which belong to an IgM function by an activation of complement system.
2. The studies on isolation and culture of human hepatocytes
(1)The yield of isolated hepatocytes obtained by pe
rfusing with a mixture of collagenase and dispase was significantly higher than that by perfusing and agitation with a collagenase solution only.
(2)The functions of cultured human hepatocytes were well preserved for 10 days in a Leibovitz L-15 medium or williams'E medium.
(3)Warm ischemia time of the liver tissue affected the yield of isolated hepatocytes. But ather liver tissue tolerated the warm ischemia for up to 120 min. The function of cultured hepatocytes was not affected by warm ischemia till 220 minutes and the hepatocytes exhibited reasonably high levels of hepatic function.
3. The effects of the artificial liver system on anhepatic xenogeneic animals
A small hybrid artificial liver module that contains 5 grams of porcine hepatocytes was manufactured. Ex vivo extracorporeal plasma perfusion studies revealed that the hybrid artificial liver which was heterogeneous to rabbits, showed a tendency to prolong the survival time of anhepatic rabbits. These results indicate that the hybrid artificial liver is a promising artificial liver support system for clinical cases. Less