Grant-in-Aid for Scientific Research (A)
|Allocation Type||Single-year Grants|
|Research Institution||KYUSHU UNIVERSITY|
FUNATSU Kazumori Kyushu Univ., Dept. Chem. Sys. Eng., Prof., 工学研究科, 教授 (80037960)
SUGIMACHI Keizo Kyushu Univ., Dept. Chem. Surgery II, Prof., 医学部, 教授 (00038762)
NAKAZAWA Kohji Kyushu Univ., Dept. Chem. Sys. Eng., Res. Assoc., 工学研究科, 助手 (00304733)
IJIMA Hiroyuki Kyushu Univ., Dept. Mater. Proc. Eng., Assoc. Prof., 工学研究科, 助教授 (10274515)
SHIRABE Ken Kyushu Univ., Dept. Chem. Surgery II, Res. Assoc., 医学部, 助手 (70264025)
SHIMADA Mitsuo Kyushu Univ., Dept.Chem. Surgery II, Lecturer, 医学部, 講師 (10216070)
宇都宮 徹 九州大学, 医学部, 助手 (30304801)
|Project Period (FY)
1998 – 1999
Completed(Fiscal Year 1999)
|Budget Amount *help
¥34,300,000 (Direct Cost : ¥34,300,000)
Fiscal Year 1999 : ¥11,300,000 (Direct Cost : ¥11,300,000)
Fiscal Year 1998 : ¥23,000,000 (Direct Cost : ¥23,000,000)
|Keywords||Hybrid artificial liver / Preclinical test / Ethics Committee / Porcine Hepatocyte / Spheroid / Polyurethane foam / ischemic liver failure pig / immunologic reaction / ヒト臨床 / 肝細胞スフェロイド / ポリウレタン発砲体 / 体外循環 / ヒト臨床応用 / 肝細胞スウェロイド / 血中アンモニア|
1. Establishment of mass isolation method of porcine hepatocytes
We established two step collagenase treatment method for mass isolation of porcine hepatocytes. This method consists of liver perfusion and cell process using collagenase solution. More than 380g porcine hepatocytes (viability : more than 85%) were isolated within 3 hours from two pig livers (570g) by this method.
2. Development of a hybrid artificial liver for children treatment
We developed a multi-capillary polyurethane foam (PUF) packed-bed module (diameter : 7.7cm, height : 20cm, volume : 1000cmィイD13ィエD1) with 100g of porcine hepatocytes for clinical application. Porcine hepatocytes in the module formed spheroids within 24 hours of culture and maintained liver specific functions at least 10days of culture. We also developed extracorporeal circulation system for an artificial liver treatment, with control of DO, pH and temperature in plasma, and refinement of circulation loop for sufficient mass transfer.
3. Preclinical tests of a hybrid artificial liver using failure pigs
As a preclinical test, we developed as artificial liver module with 200g of porcine hepatocytes, and applied to warm ischemic liver failure pigs of 25kg. Consequently, we could successfully indicate the care effect by the artificial liver in suppression of increase of blood ammonia, maintenance of blood glucose, extension of survival time and so on.
4. Study of modulation of immunologic reaction between human serum and porcine hepatocytes
When the porcine hepatocytes were cultured in human serum, immunologic reaction of human C3 against porcine hepatocytes occurred positively. We found that damage of porcine hepatocytes by human C3 could be decreased by the addition of nafamostat mesliate.
These results suggest that our device will become a promising method for clinical treatment. At present, the device have been applied to the Ethics Committee of Kyushu University for clinical application.