| Project/Area Number |
24500500
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Central Institute for Experimental Animals |
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Principal Investigator |
HARUO Hashimoto 公益財団法人実験動物中央研究所, その他部局等, 研究員 (30353478)
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| Co-Investigator(Kenkyū-buntansha) |
HIGUCHI Yuichiro 公益財団法人実験動物中央研究所, その他部局等, 研究員 (00596281)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | アデノ随伴ウィルス / ヒト化肝臓マウス / Pdx1 / 糖尿病 / Pｄｘ１ / PDX-1 |
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| Outline of Final Research Achievements |
The aim of this research was to establish a method of efficiently inducing the transdifferentiation of “human hepatocytes into human Langerhans islet or beta cells” using humanized liver mice in vivo.
First, an attempt was made to efficiently prepare adeno-associated virus (AAV) with Pdx1-GFP. During preparation of AAV, the ratio of the amount of transgene, helper vector, and capsid vector DNA μg to the amount of Lipofectamine μl was 2:6. This resulted in successful preparation of AAV2 and AAV-DJ/8 with transgene. That said, the production of AAV-DJ/8 was about 1/20th of that of AAV2. This means that AAV2 was produced efficiently.Results indicated that there was substantial transfection in areas of the “human” liver in a mouse with a humanized liver. Thus, a mouse with a humanized liver is a suitable way of assessing gene therapy using AAV, and AAV2- Pdx1-Ngn3-GFP and AAV2- Pdx1-GFP were rapidly effective as diabetes treatments, as “human” experiments in mice revealed.
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