| Project/Area Number |
15591371
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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 | National Hospital Organization Nagasaki Medical Center Clinical Research Center |
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Principal Investigator |
FUJIOKA Hikaru National Hospital Organization, Nagasaki Medical Center, Clinical Research Center, Department of Function & Morphology, Director, 臨床研究センター, 機能形態研究部長 (00264226)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIBASHI Hiromi National Hospital Organization, Nagasaki Medical Center, Clinical Research Center, Clinical Research Center, Director, 臨床研究センター, 臨床研究センター長 (80127969)
YATSUHASHI Hiroshi National Hospital Organization, Nagasaki Medical Center, Clinical Research Center, Department of Therapeutic Research, Director, 臨床研究センター, 治療研究部長 (50360855)
NAKAMURA Minoru National Hospital Organization, Nagasaki Medical Center, Clinical Research Center, Department of Advanced Medical Research, Director, 臨床研究センター, 先端技術研究部長 (40217906)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Human hepatocyte / Bank of hepatocytes / Bioartificial liver / Hepatocyte transplantation / Gene transfer / キメラマウス / 人畜共通感染症 / 凍結保存 / ヒト肝細胞化マウス / 肝不全 |
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| Research Abstract |
Liver transplantation improves the survival rate and quality of life for patients with life-threatening liver diseases. This therapy, however, is associated with several problems including significant morbidity, mortality and the shortage of liver donors. Therefore, the demand for clinical use of bioartificial liver and isolated hepatocytes. Unfortunately, the number of donor livers available for human hepatocyte isolation is limited. Other animal species have been suggested as a potential source of hepatocytes for these therapies. However, immunologic and physiologic barriers to cross-species use have yet to be overcome. In addition, there is another concern about zoonosis so that human hepatocytes shoud be used for these therapies.
The purpose of this study is to establish an efficient isolation and cryopreservation method for the human hepatocyte bank. In addition, efficiency of HVJ-liposome for gene transfer into porcine hepatocytes was investigated.
Results :
Human hepatocytes were harvested from 16 volunteers with metastatic liver tumors or hemangiomas using a modification of the two step-collagenase perfusion technique. The yield of freshly isolated hepatocytes averaged more than 8.5x10^6/liver gr.with 80% viability. The average viability of thawed hepatocytes that had been cryopreserved for 30days was 62% and plating efficiency was 40% in the solution of UW (University of Wisconsin) with human fresh frozen plasma (FFP). The use of UW solution and FFP significantly improved the viability and plating efficiency of cryopreserved human hepatocytes.
Now, we preserve 3.0x10^<10> human hepatocytes. Therefore, we can treat three patients with fatal liver failure. However, there have been no patients for the hepatocyte-based therapy during this study.
HVJ-liposome vector was safe and feasible modality for hepatocyte-directed gene transfer in pigs. It might be suitable for clinical hepatocyte-gene therapy trials.
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