| Project/Area Number |
12309003
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
広領域
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MAKUUCHI Masatoshi The University of Tokyo, Department of Surgery, Professor, 医学部附属病院, 教授 (60114641)
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| Co-Investigator(Kenkyū-buntansha) |
SAWASAKI Tohru Experimental farm of University of Agriculture, Professor, 農学部附属牧場, 教授 (00012047)
SUGAWARA Yasuhiko The University of Tokyo, Department of Surgery, Assistant Professor, 医学部附属病院, 助教授 (90313155)
NARUSE Katsutoshi The University of Tokyo, Department of Surgery, research associate, 医学部附属病院, 助手 (50291323)
SAKAI Yasuyuki The University of Tokyo, Industrial Institute for Sceince, Assistant Professor, 生産技術研究所, 助教授 (00235128)
TOJO Hideaki The University of Tokyo, Department of Agriculture, Professor, 大学院・農学生命科学研究科, 教授 (20041668)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥47,360,000 (Direct Cost: ¥40,700,000、Indirect Cost: ¥6,660,000)
Fiscal Year 2002: ¥13,000,000 (Direct Cost: ¥10,000,000、Indirect Cost: ¥3,000,000)
Fiscal Year 2001: ¥15,860,000 (Direct Cost: ¥12,200,000、Indirect Cost: ¥3,660,000)
Fiscal Year 2000: ¥18,500,000 (Direct Cost: ¥18,500,000)
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| Keywords | Bioartificial liver / Transgenic animal / Transgenic mouse / Transgenic pig / human albumin DNA / EGFP / EGFP遺伝子 / トランスジェニックマウス / トランスジェニックブタ / ハイブリッド型人工肝臓 / トランスジェニック / マイクロマニピュレーション |
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| Research Abstract |
In research on extracorporeal bioartificial liver perfusion system using xenogeneic liver, the chief purpose of the transgenic technique is to knock out genes of xenogeneic proteins, and transduce genes of human proteins into the animal. As an initial approach, we planned to transduce human albumin cDNA into porcine germ cells to produce porcine liver which can secret human albumin for the purpose of utilizing it for bioartificial liver support, we produced a gene of human albumin cDNA (hAlb) and EGFP (enhanced green fluorescent protein) gene, pCX-hAlb-EGFP. We transfected it initially into NIH3T3 cells using lipofection method, secondly into mouse germ cells by classic microinjection method, and thirdly into porcine germ cells by intracytoplasmic sperm injection (ICSI) method in order to induce expression of human albumin in the cultured cells and animal liver. As the results, NIH3T3 cells showed ultraviolet lightening and expressed human albumin in Northern hybridization analysis, and a transgenic pig was produced of which the skin showed ultraviolet lightening and integration of human albumin gene in PCR analysis. It was a pity that the pig was born dead, but this was initial epoch-making report of expression of EGFP and human albumin in TG pig produced by ICSI method.
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