Abstract
Successful gene therapy technology relies on the delivery of the therapeutic product into appropriate target cells. Gene delivery to mesenchymal stem cells (MSCs) has been proposed as a mechanism to promote the augmentation of tissue-engineered replacement systems. In particular, MSCs are attractive targets for gene delivery systems, because they can differentiate, in response to various molecular signals, into many types of committed cells. Introduction of transgene of interest into autologous stem cell types poses an attractive cell-based delivery strategy. MSCs divide rapidly and, because of their high amphotropic receptor levels, are readily transducible with integrating vectors and maintain transgene expression in vitro and in vivo without affecting multipotentiality. The unique biology of MSCs predetermines them to become valuable cytoreagents for gene therapy approaches in future. This chapter describes methods and associated materials for transducing mesenchymal stem cells with a desired nucleic acid.
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References
1. Krause, D. S., et al., (2001) Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell. Cell, 105(3): 369–377.
2. Morizono, K., et al., (2003) Multilineage cells from adipose tissue as gene delivery vehicles. Hum. Gene Ther. 14(1):59–66.
3. Hamada, H., et al., (2005) Mesenchymal stem cells (MSC) as therapeutic cytoreagents for gene therapy. Cancer Sci. 96(3): 149–156.
4. Scherr, M. and Eder, M., (2002) Gene transfer into hematopoietic stem cells using lentiviral vectors. Curr. Gene Ther. 2(1): 45–55.
5. Davis, B. M., Humeau, L., and Dropulic, B. (2004) In vivo selection for human and murine hematopoietic cells transduced with a therapeutic MGMT lentiviral vector that inhibits HIV replication. Mol. Ther. 9(2): 160–172.
6. Haviernik, P. and Bunting, K. D. (2004) Safety concerns related to hematopoietic stem cell gene transfer using retroviral vectors. Curr. Gene Ther. 4(3): 263–276.
7. Colter, D. C., et al., (2000) Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc. Natl Acad. Sci. U S A. 97(7): 3213–3218.
8. Izadpanah, R., et al., (2005) Characterization of multipotent mesenchymal stem cells from the bone marrow of rhesus macaques. Stem Cells Dev. 14(4): 440–451.
9. Friedenstein, A. J., (1976) Precursor cells of mechanocytes. Int. Rev. Cytol. 47: 327–359.
10. Howlett, C. R., et al., (1986) Mineralization in in vitro cultures of rabbit marrow stromal cells. Clin. Orthop. 213: 251–263.
11. Lackner, A. A., et al., (1988) Distribution of a macaque immunosuppressive type D retrovirus in neural, lymphoid, and salivary tissues. J. Virol. 62(6): 2134–2142.
12. Kanazawa, T., et al., (2001) Gamma-rays enhance rAAV-mediated transgene expression and cytocidal effect of AAV-HSVtk/ganciclovir on cancer cells. Cancer Gene Ther. 8(2): 99–106.
13. Pittenger, M. F., et al., (1999) Multilineage potential of adult human mesenchymal stem cells. Science. 284(5411): 143–147.
14. Mezey, E., et al., (2000) Turning blood into brain: cells bearing neuronal antigens generated in vivo from bone marrow. Science. 290(5497): 1779–1782.
15. Orlic, D., (2003) Adult bone marrow stem cells regenerate myocardium in ischemic heart disease. Ann. N Y Acad. Sci. 996: 152–157.
16. Ferrari, G., et al., (1998) Muscle regeneration by bone marrow-derived myogenic progenitors. Science. 279(5356): 1528–1530.
17. Theise, N. D., et al., (2000) Liver from bone marrow in humans. Hepatology. 32(1): 11–16.
18. Eglitis, M. A. and Mezey, E. (1997) Hematopoietic cells differentiate into both microglia and macroglia in the brains of adult mice. Proc. Natl Acad. Sci. U S A. 94(8): 4080–4085.
19. Kascsak, R. J., et al., (1982) Virological studies in amyotrophic lateral sclerosis. Muscle Nerve. 5(2): 93–101.
20. Zuk, P.A., et al., (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 7(2): 211–228.
21. Zuk, P. A., et al., (2002) Human adipose tissue is a source of multipotent stem cells. Mol. Biol. Cell. 13(12): 4279–4295.
22. Mizuno, H., et al., (2002) Myogenic differentiation by human processed lipoaspirate cells. Plast. Reconstr. Surg. 109(1): 199–209; discussion 210–211.
23. Abdallah, B., Sachs, L., and Demeneix, B. A. (1995) Non-viral gene transfer: applications in developmental biology and gene therapy. Biol. Cell. 85(1): 1–7.
Bosch, P., Pratt, S. L. and Stice, S. L. Isolation, characterization, gene modification and nuclear reprogramming of porcine mesenchymal stem cells. Biol Reprod. 2005.
25. Conget, P. A., Allers, C. and Minguell, J.J. (2001) Identification of a discrete population of human bone marrow-derived mesenchymal cells exhibiting properties of uncommitted progenitors. J. Hematother. Stem Cell Res. 10(6): 749–758.
26. Kumar, S., et al., (2004) Osteogenic differentiation of recombinant adeno-associated virus 2-transduced murine mesenchymal stem cells and development of an immunocompetent mouse model for ex vivo osteoporosis gene therapy. Hum. Gene Ther. 15(12): 1197–1206.
27. Lee, K., et al., (2001) Human mesenchymal stem cells maintain transgene expression during expansion and differentiation. Mol. Ther. 3(6): 857–866.
28. Lu, L., et al., (2005) Therapeutic benefit of TH-engineered mesenchymal stem cells for Parkinson's disease. Brain Res. Brain Res. Protoc. 15(1): 46–51.
29. Lu, S., et al., (2005) (Construction of an adeno-associated virus vector expressing CTLA-4Ig and its expression in the transplanted liver allografts). Zhonghua Gan Zang Bing Za Zhi, 13(3): 183–186.
30. Van Damme, A., et al., (2004) Onco-retroviral and lentiviral vector-based gene therapy for hemophilia: preclinical studies. Semin. Thromb. Hemost. 30(2): 185–195.
31. Zhang, X.Y., La Russa, V.F. and Reiser, J. (2004) Transduction of bone-marrow-derived mes-enchymal stem cells by using lentivirus vectors pseudotyped with modified RD114 envelope glycoproteins. J. Virol. 78(3): 1219–1229.
32. Naldini, L., (1998) Lentiviruses as gene transfer agents for delivery to non-dividing cells. Curr. Opin. Biotechnol. 9(5): 457–463.
33. Naldini, L., et al., (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science. 272(5259): 263–267.
34. Miyoshi, H., et al., (1998) Development of a self-inactivating lentivirus vector. J Virol. 72(10): 8150–8157.
35. Zhang, B., et al., (2004) The significance of controlled conditions in lentiviral vector titration and in the use of multiplicity of infection (MOI) for predicting gene transfer events. Genet. Vaccines Ther. 2(1): 6.
36. Skipski, V.P., Smolowe, A.F. and Barclay, M. (1967) Separation of neutral glycosphingolipids and sulfatides by thin-layer chromatography. J. Lipid Res. 8(4): 295–299.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Izadpanah, R., Bunnell, B.A. (2008). Gene Delivery to Mesenchymal Stem Cells. In: Prockop, D.J., Bunnell, B.A., Phinney, D.G. (eds) Mesenchymal Stem Cells. Methods in Molecular Biology™, vol 449. Humana Press. https://doi.org/10.1007/978-1-60327-169-1_11
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DOI: https://doi.org/10.1007/978-1-60327-169-1_11
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