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Blood, 15 May 2004, Vol. 103, No. 10, pp. 3710-3716. Prepublished online as a Blood First Edition Paper on January 22, 2004; DOI 10.1182/blood-2003-07-2414. This Article Full Text Full Text (PDF) All Versions of this Article: 2003-07-2414v1 103/10/3710    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Horn, P. A. Articles by Kiem, H.-P. Search for Related Content PubMed PubMed Citation Articles by Horn, P. A. Articles by Kiem, H.-P. Related Collections Hematopoiesis and Stem Cells Gene Expression Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Efficient lentiviral gene transfer to canine repopulating cells using an overnight transduction protocol Peter A. Horn, Kirsten A. Keyser, Laura J. Peterson, Tobias Neff, Bobbie M. Thomasson, Jesse Thompson, and Hans-Peter Kiem From the Clinical Division, Fred Hutchinson Cancer Research Center, Seattle WA; and the Department of Medicine, University of Washington School of Medicine, Seattle WA. The use of lentiviral vectors for the transduction of hematopoietic stem cells has evoked much interest owing to their ability to stably integrate into the genome of nondividing cells. However, published large animal studies have reported highly variable gene transfer rates of typically less than 1%. Here we report the use of lentiviral vectors for the transduction of canine CD34+ hematopoietic repopulating cells using a very short, 18-hour transduction protocol. We compared lentiviral transduction of hematopoietic repopulating cells from either stem cell factor (SCF)– and granulocyte-colony stimulating factor (G-CSF)–primed marrow or mobilized peripheral blood in a competitive repopulation assay in 3 dogs. All dogs engrafted rapidly within 9 days. Transgene expression was detected in all lineages (B cells, T cells, granulocytes, and red blood cells as well as platelets) indicating multilineage engraftment of transduced cells, with overall long-term marking levels of up to 12%. Gene transfer levels in mobilized peripheral blood cells were slightly higher than in primed marrow cells. In conclusion, we show efficient lentiviral transduction of canine repopulating cells using an overnight transduction protocol. These results have important implications for the design of stem cell gene therapy protocols, especially for those diseases in which the maintenance of stem cells in culture is a major limitation. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: G. D. Trobridge, B. C. Beard, C. Gooch, M. Wohlfahrt, P. Olsen, J. Fletcher, P. Malik, and H.-P. Kiem Efficient transduction of pigtailed macaque hematopoietic repopulating cells with HIV-based lentiviral vectors Blood, June 15, 2008; 111(12): 5537 - 5543. [Abstract] [Full Text] [PDF] K. Aoyama, C. Delaney, B. Varnum-Finney, A. D. Kohn, R. T. Moon, and I. D. Bernstein The Interaction of the Wnt and Notch Pathways Modulates Natural Killer Versus T Cell Differentiation Stem Cells, October 1, 2007; 25(10): 2488 - 2497. [Abstract] [Full Text] [PDF] J. L. Gori, K. Podetz-Pedersen, D. Swanson, A. D. Karlen, R. Gunther, N. V. Somia, and R. S. McIvor Protection of Mice from Methotrexate Toxicity by ex Vivo Transduction Using Lentivirus Vectors Expressing Drug-Resistant Dihydrofolate Reductase J. Pharmacol. Exp. Ther., September 1, 2007; 322(3): 989 - 997. [Abstract] [Full Text] [PDF] X.-B. Zhang, J. L. Schwartz, R. K. Humphries, and H.-P. Kiem Effects of HOXB4 Overexpression on Ex Vivo Expansion and Immortalization of Hematopoietic Cells from Different Species Stem Cells, August 1, 2007; 25(8): 2074 - 2081. [Abstract] [Full Text] [PDF] H.-P. Kiem, J. Allen, G. Trobridge, E. Olson, K. Keyser, L. Peterson, and D. W. Russell Foamy virus-mediated gene transfer to canine repopulating cells Blood, January 1, 2007; 109(1): 65 - 70. [Abstract] [Full Text] [PDF] T. R. Bauer Jr, M. Hai, L. M. Tuschong, T. H. Burkholder, Y.-c. Gu, R. A. Sokolic, C. Ferguson, C. E. Dunbar, and D. D. Hickstein Correction of the disease phenotype in canine leukocyte adhesion deficiency using ex vivo hematopoietic stem cell gene therapy Blood, November 15, 2006; 108(10): 3313 - 3320. [Abstract] [Full Text] [PDF] T. Yoshioka, N. Ageyama, H. Shibata, T. Yasu, Y. Misawa, K. Takeuchi, K. Matsui, K. Yamamoto, K. Terao, K. Shimada, et al. Repair of Infarcted Myocardium Mediated by Transplanted Bone Marrow-Derived CD34+ Stem Cells in a Nonhuman Primate Model Stem Cells, March 1, 2005; 23(3): 355 - 364. [Abstract] [Full Text] [PDF]

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