| Project/Area Number |
12671134
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General surgery
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| Research Institution | Asahikawa Medical College |
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Principal Investigator |
OBARA Mitsuhiro Asahikawa Medical College, Assistant, 医学部, 助手 (00214221)
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| Co-Investigator(Kenkyū-buntansha) |
TOMITA Ichirou Asahikawa Medical College, Assistant, 医学部, 医員 (00332487)
澤 雅之 旭川医科大学, 医学部, 助手 (70226059)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Hepatocyte transplantation / Non-heart-beating cadaver / 肝細胞分離 / 細胞移植 |
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| Research Abstract |
1. Development of isolation technique for hepatocyte from non-heart-beating cadaver
(1) Trans-portal perfusion method
Insufficient heparinization leads to much residual blood in the livers and poor yield of viable hepatocytes. Viability (yield: viable hepatocyte/wet liver weight) of isolated hepatocytes at 0, 30, 60, 90, 120 min of warm ischemia is 92.0% (1.2×10^7/wet liver), 70.2 (8.1×10^5), 43.2 (5.6×10^3), 12.3 (1.5×10^2), 1.4(1.7×10), respectively.
(2) Multipuncture perfusion method
After multipuncture and perfusion of collagenase solution, hepatocytes could not be isolated and still remained in clusters.
2. Purification of isolated hepatocyte
No viable isolated hepatocyte was obtained after purification of 30 min warm ischemic cells by the use of Percoll. No purified 30 min warm ischemic hepatocytes were cultured in MEM. Membrane of hepatocytes showed budding formation and urea synthesis of hepatocytes from addition of NH_4Cl deteriorated.
DcAMP and Chlorpromazine have no good effect for isolation of hepatocytes from non-heart-beating cadaver. It is necessary for hepatocyte transplantation from non-heart-beating cadaver to improve isolation technique.
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