Summary
Adult bone marrow stem cell is an ideal target for gene therapy of genetic diseases, selected malignant diseases, and AIDS. The in vivo approach of lentivirus vector (LV)-mediated stem cell gene transfer by intrafemoral (IF) injection can take full advantage of any source of stem cells residing in the bone cavity. Such an approach may avoid several difficulties encountered by ex vivo hematopoietic stem cell (HSC) gene transfer. We have shown that both HSC and mesenchymal stem/progenitor cells (MSC) can be genetically modified successfully by a single “in situ” IF injection in their natural “niche” in mice without any preconditioning. This approach may provide a novel application for treatment of human diseases, and represent an interesting new tool to study adult stem cell plasticity and the nature of unperturbed hematopoiesis.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cheshier, S.H., S.J. Morrison, X. Liao, and I.L. Weissman, In vivo proliferation and cell cycle kinetics of long-term self-renewing hemat-opoietic stem cells. Proc Natl Acad Sci U S A, 1999. 96 (6) : pp. 3120–3125
Pan, D., R. Gunther, W. Duan, S. Wendell, W. Kaemmerer, T. Kafri, I.M. Verma, and C.B. Whitley, Biodistribution and toxicity studies of VSVG-pseudotyped lentiviral vector after intravenous administration in mice with the observation of in vivo transduction of bone marrow. Mol Ther, 2002. 6 (1) : pp. 19–29
Follenzi, A., G. Sabatino, A. Lombardo, C. Boccaccio, and L. Naldini, Efficient gene delivery and targeted expression to hepatocytes in vivo by improved lentiviral vectors. Hum Gene Ther, 2002. 13 (2) : pp. 243–260
McCauslin, C.S., J. Wine, L. Cheng, K.D. Klarmann, F. Candotti, P.A. Clausen, S.E. Spence, and J.R. Keller, In vivo retroviral gene transfer by direct intrafemoral injection results in correction of the SCID phenotype in Jak3 knock-out animals. Blood, 2003. 102 (3) : pp. 843–848
Worsham, D.N., T. Schuesler, C. von Kalle, and D. Pan, In vivo gene transfer into adult stem cells in unconditioned mice by in situ delivery of a lentiviral vector. Mol Ther, 2006. 14 (4) : pp. 514–524
Jiang, Y., B.N. Jahagirdar, R.L. Reinhardt, R.E. Schwartz, C.D. Keene, X.R. Ortiz-Gonzalez, M. Reyes, T. Lenvik, T. Lund, M. Blackstad, J. Du, S. Aldrich, A. Lisberg, W.C. Low, D.A. Largaespada, and C.M. Verfaillie, Pluripotency of mesenchymal stem cells derived from adult marrow Nature, 2002. 418 (6893) : pp. 41–49
Herzog, E.L., L. Chai, and D.S. Krause, Plasticity of marrow derived stem cells. Blood, 2003. 102 (10) : pp. 3483–3493
Schnitzlein, W.M. and M.E. Reichmann, Characterization of New Jersey vesicular stomatitis virus isolates from horses and black flies during the 1982 outbreak in Colorado. Virology, 1985. 142 (2) : pp. 426–431
Burns, J.C., T. Friedmann, W. Driever, M. Burrascano, and J.K. Yee, Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells. Proc Natl Acad Sci U S A, 1993. 90 (17) : pp. 8033–8037
Naldini, L., U. Blomer, P. Gallay, D. Ory, R. Mulligan, F.H. Gage, I.M. Verma, and D. Trono, In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science, 1996. 272 (5259) : pp. 263–267
Poeschla, E.M., F. Wong-Staal, and D.J. Looney, Efficient transduction of nondividing human cells by feline immunodeficiency virus lentiviral vectors. Nat Med, 1998. 4 (3) : pp. 354–357
Mazurier, F., M. Doedens, O.I. Gan, and J.E. Dick, Rapid myeloerythroid repopulation after intrafemoral transplantation of NOD-SCID mice reveals a new class of human stem cells. Nat Med, 2003. 9 (7) : pp. 959–963
Acknowledgments
The author would like to thank the Veterinary Service at Cincinnati Children's Hospital Medical Center for their support and consultation. This work was supported in part by Translational Research Initiative grant from Cincinnati Children's Foundation, and the National Institutes of Health (AI061703).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press
About this protocol
Cite this protocol
Pan, D. (2009). In Situ (In Vivo) Gene Transfer into Murine Bone Marrow Stem Cells. In: Baum, C. (eds) Genetic Modification of Hematopoietic Stem Cells. Methods In Molecular Biology™, vol 506. Humana Press. https://doi.org/10.1007/978-1-59745-409-4_12
Download citation
DOI: https://doi.org/10.1007/978-1-59745-409-4_12
Publisher Name: Humana Press
Print ISBN: 978-1-58829-980-2
Online ISBN: 978-1-59745-409-4
eBook Packages: Springer Protocols