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Genetic manipulation and transplantation of cynomolgus embryonic stem cells

Research Project

Project/Area Number 13671080
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionJichi Medical School

Principal Investigator

HANAZONO Yutaka  M.D., Ph.D., Assistant Professor, Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, 医学部, 講師 (70251246)

Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordsembryonic stem cells / lentiviral vector / cynomolgus monkeys / primate model / NOD / SCID / 胎仔移植 / カニクイザル / ヒツジ / GFP / OP9 / BMP-4 / ヒツジ胎仔
Research Abstract

The ability to stably introduce genetic material into primate embryonic stem (ES) cells could allow broader application. In this study, we examined lentiviral gene transfer into cynomolgus ES cells. When cynomolgus ES cells were transduced once with a simian immunodeficiency virus (SIV)-based lentivirus vector encoding the green fluorescent protein (GFP) gene, most cells (around 90%) fluoresced and high levels of GFP expression persisted for 5 months without selection procedures. In addition, high levels of GFP expression were observed during embryoid body formation. On the other hand, transduction of mouse ES cells with the SIV based vector resulted in lower gene transfer rates, implying that primate viral vectors can transduce primate ES cells more efficiently than mouse ES cells. The use of GFP as a reporter gene allows direct and simple detection of successfully transduced ES cells and facilitates monitoring of ES cell proliferation and differentiation both in vitro and potentially in vivo. Furthermore, this highly efficient gene transfer method allows faithful gene delivery to primate ES cells with potential for both research and therapeutic application.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Nagashima, T. et al.: "In vivo expansion of gene-modified hematopoietic cells by a novel selective amplifier gene utilizing the erythropoietin receptor as a molecular switch"Biochem. Biophys. Res. Comm.. (発表予定). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono, Y. et al.: "Genetic manipulation of primate embryonic and hematopoietic stem cells with simian lentivirus vectors"Trends in Cardiovascular Medicine. 13巻. 106-110 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ageyama, N. et al.: "Safe and efficient methods of autologous hematopoietic stem cell transplantation for biomedical research in cynomolgus monkeys"Comp. Med.. 52巻. 445-451 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono, Y. et al.: "Introduction of the GFP gene into hematopoietic stem cells results in prolonged discrepancy of in vivo transduction levels between bone marrow progenitors and peripheral blood cells in nonhuman primates"J. Gene Med.. 4巻. 470-477 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Asano, T. et al.: "Highly efficient gene transfer into primate embryonic stem cells with a simian lentivirus vector"Mol. Ther.. 6巻. 162-168 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono, Y. et al.: "In vivo selective expansion of gene-modified hematopoietic cells in a nonhuman primate model"Gene Ther.. 9巻. 1055-1064 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Muramatsu, M. et al.: "Reversible integration of the dominant negative retinoid receptor gene for ex vivo expansion of hematopoietic stem/progenitor cells"Biochem. Biophys. Res. Commun.. 285巻. 891-896 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono, Y. et al.: "Gene transfer into nonhuman primate hematopoietic stem cells : implications for gene therapy"Stem Cells. 19巻. 12-23 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nagashima,T., et al.: "In vivo expansion of gene-modified hematopoietic cells by a novel selective amplifier gene utilizing the erythropoietin receptor as a molecular switch"Biochem.Biophys.Res.Comm.. in press.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono Y, et al.: "Genetic manipulation of primate embryonic and hematopoietic stem cells with simian lentivirus vectors"Trends in Cardiovascular Medicine. 13. 106-110 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ageyama,N., et al.: "Safe and efficient methods of autologous hematopoietic stem cell transplantation for biomedical research in cynomolgus monkeys"Comp.Med.. 52. 445-451 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono,Y., et al.: "Introduction of the GFP gene into hematopoietic stem cells results in prolonged discrepancy of in vivo transduction levels betweem bone marrow progenitors and peripheral blood cells in nonhuman primates"J.Gene Med.. 4. 470-477 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Asano,T., et al.: "Highly efficient gene transfer into primate embryonic stem cells with a simian lentivirus vector"Mol.Ther.. 6. 162-168 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono,Y., et al.: "In vivo selective expansion of gene-modified hematopoietic cells in a nonhuman primate model"Gene Ther.. 9. 1055-1064 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Muramatsu,M., Hanazono,Y., Ogasawara,Y., Okada,T., Mizukami,H., Kume,A., Mizoguchi,H., and Ozawa,K.: "Reversible integration of the dominant negative retinoid receptor gene for ex vivo expansion of hematopoietic stem/progenitor cells"Biochem.Biophys.Res.Commun.. 285. 891-896 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono,Y., Terao,K., and Ozawa,K.: "Gene transfer into nonhuman primate hematopoietic stem cells : implications for gene therapy"Stem Cells. 19. 12-23 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hanazono, Y.et al.: "Genetic manipulation of primate embryonic and hematopoietic stem cells with simian lentivirus vectors"Trends in Cardiovascular Medicine. (発表予定). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kume, A.et al.: "In vivo expansion of transduced murine hematopoietic cells with a selective amplifier gene"J. Gene Med.. (発表予定). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ageyama, N. et al.: "Safe and efficient methods for cynomolgus monkey autologous hematopoietic stem cell transplantation for biomedical researches"Comp. Med.. 52巻. 445-451 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kume, A. et al.: "Selective expansion of transduced cells for hematopoietic stem cell gene therapy"Int. J. Hematol.. 76巻. 299-304 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Okada, T. et al.: "AAV mediated gene transfer for gene therapy of ischemia-induced neuronal death"Methods. Enzymol.. 346巻. 378-393 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Wang, L. et al.: "Delayed delivery of AAV-GDNF prevents nigral neurodegeneration and promotes functional recovery in a rat model of Parkinson's disease"Gene Ther.. 9巻. 381-389 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Muramatsu, M. et al.: "Reversible integration of the dominant negative retinoid receptor gene for ex vivo expansion of hematopoietic stem/progenitor cells"Biochem. Biophys. Res. Commun.. 285巻. 891-896 (2001)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hanazono, Y. et al.: "Gene transfer into nonhuman primate hematopoietic stem cells : implications for gene therapy"Stem Cells. 19巻. 12-23 (2001)

    • Related Report
      2002 Annual Research Report
  • [Publications] Muramatsu, M., et al.: "Reversible integration of the dominant negative retinoid receptor gene for ex vivo expansion of hematopoietic stem/progenitor cells"Biochem. Biophys. Res. Commun.. 285巻. 891-896 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kang, E.M.et al.: "Persistent low-level engraftment of rhesus peripheral blood progenitor cells transduced with the Fanconi anemia C gene after conditioning with low-dose irradiation"Mol. Ther.. 3巻. 911-919 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hanazono, Y. et al.: "Gene transfer into nonhuman primate hematopoietic stem cells : implications for gene therapy"Stem Cells. 19巻. 12-23 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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