| Project/Area Number |
59890011
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
広領域
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| Research Institution | Kitasato University |
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Principal Investigator |
YOSHIZATO Katsutoshi Dept.of Plastic Surgery,Kitasato Univ. School of Med., 医学部, 助教授 (20095516)
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| Co-Investigator(Kenkyū-buntansha) |
NAGAYOSHI Kumiko Ultrastruc.Res.Div.,Kitasato Uni. School of Nursing., 看護学部, 助手 (00189169)
NAMIKI Hideo Dept.of Biology, School of Education, Waseda Univ., 教育学部, 助手 (50120928)
KUSUNOKI Shinichiro Institute of Life Science, Advance Co. Ltd., 生命科学研究所, 所長
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| Project Period (FY) |
1984 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1986: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1985: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1984: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Keywords | Artificial Liver / Collagen / Hepatocytes / Fibroblasts / コラーゲン / 細胞組込み型人工臓器 / 酸素分圧 |
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| Research Abstract |
The liver of mammals plays many important functions as a center of metabolic processing of all materials taken up by the animal. This fact does not allow us make a liver substitute for the person who had severe demages to this organ. This situation of the liver substitute contrasts to that of a heart or renal substitute. The only way to make an artificial liver is to utilize hepatocytes as a functional key unit. For this aim, we developed a way for culturing hepatocytes in a healthy condition for a long period of time. We sought for the optimal condition for hepatocytes cultures from the following points of view. 1. Introduction of extracellular matrices (ECM) as a culture substrate. 2. Optimal gas phase. 3. Interactions of hepatocytes with non-parenchymal cells. 4. Development of perfusion culture 5. Development of three-dimensional culture. 1. ECM. 1.1. Collagen. The rate of DNA synthesis of hepatocytes was surpressed markedly when cultured on collagen fibrils. However, the secretion of albumin was enhanced on the same substrates. 1.2. Separation of liver matrix. ECM of the liver was extracted by perfusing the liver with urea. This extract was found to be an effective substrate for hepatocytes. 2. Gas phase of the hepatocyte cultures. Hepatocytes required heperoxia (40-50% in partial presure) for the optimal adhesion, spreading and viability in culture. 3. Requirement of fibroblasts for a long term culture. When hepatocytes were co-cultured with fibroblasts (3T3 or lung fibroblasts), they were maintained in morphologically and functionally healthy conditions over a term of 40 days. 4. Perfusion culture. We developed a culture equipment, where hepatocytes can be maintained in a perfused media, controlling pH and oxygen concentrations at a desired level. 5. Three-dimensional culture. Hepatocytes were cultured three-dimensionally in a matrix of collagen gels. This technique provides us a method to immobilize hepatocytes in a compact mass.
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