Grant-in-Aid for Scientific Research (A).
|Research Institution||KYOTO UNIVERSITY|
YAMAOKA Yoshiro KYOTO UNIVERSITY, Graduate School of Medicine, Professor, 医学研究科, 教授 (90089102)
西河 芳樹 ノバルティスファーマ株式会社, 取締役研究開発本部長
山本 孝史 農林水産省, 家畜衛生試験場, 部長
山内 清明 京都大学, 大学院・医学研究科, 助手 (00291427)
IWAO Ikai KYOTO UNIVERSITY, Graduate School of Medicine, Lecturer, 医学研究科, 講師 (60263084)
TERAJIMA Hiroaki KYOTO UNIVERSITY, Graduate School of Medicine, Assistant, 医学研究科, 助手 (40314215)
YAMAMOTO Koshi National institute of Animal Health
NISHIKAWA Yoshiki Novartis Pharma K. K.
|Project Fiscal Year
1998 – 2000
Completed(Fiscal Year 2001)
|Budget Amount *help
¥31,300,000 (Direct Cost : ¥31,300,000)
Fiscal Year 2000 : ¥10,300,000 (Direct Cost : ¥10,300,000)
Fiscal Year 1999 : ¥10,000,000 (Direct Cost : ¥10,000,000)
Fiscal Year 1998 : ¥11,000,000 (Direct Cost : ¥11,000,000)
|Keywords||liver assist device / extracorperal live perfusion / xenogeneic liver / baboon / DAF / transgenic pig / PREV / 人工肝臓 / 体外肝灌流 / 異種肝臓 / ヒヒ / 遺伝子導入ブタ / ブタ内因性レトロウイルス / 体外異種肝灌流 / レトロウィルス / IgY / ブタ / レトロウイルス|
The aim of this study was to detect and analyze risk factors of direct cross-circulation between porcine liver and non-human primates prior to a clinical application of extracorporeal liver per fusion as a liver assist method.
1) We established the optimized condition for ling-term xenogeneic extracorporeal liver perfusion. The survival of model animals induced acute liver failure prolonged by direct cross-circulation with this system.
2) Healthy baboons were directly connected to the ECLP system. When cross-circulation was continued for 10 hours, severe macroscopic hemolysis occurred and the baboon died of acute renal failure two days later. The macroscopic hemolysis should be attributed to direct erythrocyte fragmentation resulting from humoral factor-mediated endothelial cell injury in the porcine liver graft. However, cross-circulation for less than 6 hours did not induced hemolysis, and baboons survived for more than one year without any organ dysfunction.
3) When direct cross-circul
ation between baboons and ECLP using human decay accelerating factor (hDAF) transgenic pig livers, little hemolysis occurred, allowing a cross-circulation for 21 +/- 5.8 hours (n=5). At the end of these experiments, all baboons were alive and did not show any serious complications. Through the perfusion period, bile production of the pig livers wag preserved in all cases.
4) In order to evaluate the risk of porcine endogenous retrovirus (PERV) infection, we observed baboons clinically and genetically after ECLP treatment using hDAF pig livers for 6 months after an exposure to the pig organ. The PERV sequences and the pig-specific centromeric sequences were detected in the peripheral blood of baboons during the early time of ECLP, but both disappeared within 1 week. No PERV or centromeric sequences were detected at 1 month, 3 months and 6 months after ECLP. There was no emergence of pig-cell microchimerism, and PERV did not infect.
These results indicate that an ECLP using an hDAF transgenic pig liver gives good performance, and shows promise for use in a liver assist device. Less