Genetic manipulation and transplantation of cynomolgus embryonic stem cells

2002 Fiscal Year Final Research Report Summary

• Project/Area Number: 13671080
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hematology
• Research Institution: Jichi 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
• Keywords: embryonic stem cells / lentiviral vector / cynomolgus monkeys / primate model / NOD / SCID / 胎仔移植 / カニクイザル / ヒツジ

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.

Research Products

• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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