2001 Fiscal Year Final Research Report Summary

Hepatocyte Transplantation from non-heart-beating cadaver

Research Project

Project/Area Number	12671134
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	General surgery
Research Institution	Asahikawa Medical College
Principal Investigator	OBARA Mitsuhiro Asahikawa Medical College, Assistant, 医学部, 助手 (00214221)
Co-Investigator(Kenkyū-buntansha)	TOMITA Ichirou Asahikawa Medical College, Assistant, 医学部, 医員 (00332487)
Project Period (FY)	2000 – 2001
Keywords	Hepatocyte transplantation / Non-heart-beating cadaver
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.