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Adenoviral Transduction of Mesenchymal Stem Cells

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Stem Cell Assays

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 407))

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Intact or genetically manipulated mesnechymal stem cells (MSCs) are being considered an important cell source for developing human cell-based therapeutic approaches. For applications in which transient, high-level expression of the transgene is necessary, adenovirus vectors have become increasingly popular gene-transfer vehicles. However, host range and cell-type tropism restrict the use of specific adenovectors, sometimes necessitating the lengthy development of vectors with appropriate cell specificity. Here, we present a versatile and inexpensive porcine MSC transduction procedure that can also be used on other cell types from various species, including human that are otherwise refractory to adenovirus infection.

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References

  1. Bosch, P., Pratt, S. L., and Stice, S. L. (2006) Isolation, characterization, gene modification and nuclear reprogramming of porcine mesenchymal stem cells. Biol Reprod 74, 46–57.

    Article  CAS  PubMed  Google Scholar 

  2. Pittenger, M. F., Mackay, A. M., Beck, S. C., Jaiswal, R. K., Douglas, R., Mosca, J. D., Moorman, M. A., Simonetti, D. W., Craig, S., and Marshak, D. R. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284: 143–7.

    Article  CAS  PubMed  Google Scholar 

  3. Woodbury, D., Schwarz, E. J., Prockop, D. J., and Black, I. B. (2000) Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 61, 364–70.

    Article  CAS  PubMed  Google Scholar 

  4. Nakamizo, A., Marini, F., Amano, T., Khan, A., Studeny, M., Gumin, J., Chen, J., Hentschel, S., Vecil, G., Dembinski, J., Andreeff, M., and Lang, F. F. (2005) Human bone marrow-derived mesenchymal stem cells in the treatment of gliomas. Cancer Res 65, 3307–18.

    CAS  PubMed  Google Scholar 

  5. Studeny, M., Marini, F. C., Champlin, R. E., Zompetta, C., Fidler, I. J., and Andreeff, M. (2002) Bone marrow-derived mesenchymal stem cells as vehicles for interferon-beta delivery into tumors. Cancer Res 62, 3603–8.

    CAS  PubMed  Google Scholar 

  6. Studeny, M., Marini, F. C., Dembinski, J. L., Zompetta, C., Cabreira-Hansen, M., Bekele, B. N., Champlin, R. E., and Andreeff, M. (2004) Mesenchymal stem cells: potential precursors for tumor stroma and targeted-delivery vehicles for anticancer agents. J Natl Cancer Inst 96: 1593–603.

    Article  CAS  PubMed  Google Scholar 

  7. Amalfitano, A. (2004) Utilization of adenovirus vectors for multiple gene transfer applications. Methods 33, 173–8.

    Article  CAS  PubMed  Google Scholar 

  8. Thoma, S. J., Lamping, C. P., and Ziegler, B. L. (1994) Phenotype analysis of hematopoietic CD34+ cell populations derived from human umbilical cord blood using flow cytometry and cDNA-polymerase chain reaction. Blood 83, 2103–14.

    CAS  PubMed  Google Scholar 

  9. Rebel, V. I., Hartnett, S., Denham, J., Chan, M., Finberg, R., and Sieff, C. A. (2000) Maturation and lineage-specific expression of the coxsackie and adenovirus receptor in hematopoietic cells. Stem Cells 18, 176–82.

    Article  CAS  PubMed  Google Scholar 

  10. Olmsted-Davis, E. A., Gugala, Z., Gannon, F. H., Yotnda, P., McAlhany, R. E., Lindsey, R. W., and Davis, A. R. (2002) Use of a chimeric adenovirus vector enhances BMP2 production and bone formation. Hum Gene Ther 13, 1337–47.

    Article  CAS  PubMed  Google Scholar 

  11. Kawabata, K., Sakurai, F., Koizumi, N., Hayakawa, T., and Mizuguchi, H. (2006) Adenovirus vector-mediated gene transfer into stem cells. Mol Pharm 3, 95–103.

    Article  CAS  PubMed  Google Scholar 

  12. Yotnda, P., Onishi, H., Heslop, H. E., Shayakhmetov, D., Lieber, A., Brenner, M., and Davis, A. (2001) Efficient infection of primitive hematopoietic stem cells by modified adenovirus. Gene Ther 8, 930–7.

    Article  CAS  PubMed  Google Scholar 

  13. Dodds, E., Piper, T. A., Murphy, S. J., and Dickson, G. (1999) Cationic lipids and polymers are able to enhance adenoviral infection of cultured mouse myotubes. J Neurochem 72, 2105–12.

    Article  CAS  PubMed  Google Scholar 

  14. Byk, T., Haddada, H., Vainchenker, W., and Louache, F. (1998) Lipofectamine and related cationic lipids strongly improve adenoviral infection efficiency of primitive human hematopoietic cells. Hum Gene Ther 9, 2493–502.

    Article  CAS  PubMed  Google Scholar 

  15. Bosch, P., Fouletier-Dilling, C., Olmsted-Davis, E. A., Davis, A. R., and Stice, S. L. (2006) Efficient adenoviral-mediated gene delivery into porcine mesenchymal stem cells. Mol Reprod Dev 73: 1393–403.

    Article  CAS  PubMed  Google Scholar 

  16. Fouletier-Dilling, C. M., Bosch, P., Davis, A. R., Shafer, J. A., Stice, S. L., Gugala, Z., Gannon, F. H., and Olmsted-Davis, E. A. (2005) Novel compound enables high-level adenovirus transduction in the absence of an adenovirus-specific receptor. Hum Gene Ther 16, 1287–97.

    Article  CAS  PubMed  Google Scholar 

  17. Olmsted, E. A., Blum, J. S., Rill, D., Yotnda, P., Gugala, Z., Lindsey, R. W., and Davis, A. R. (2001) Adenovirus-mediated BMP2 expression in human bone marrow stromal cells. J Cell Biochem 82, 11–21.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Julie Nelson from the Center for Tropical and Emerging Global Diseases, Flow Cytometry Facility at The University of Georgia for her assistance with flow cytometry analysis and funding from ViaGen.

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Authors and Affiliations

  1. Departamento de Biología Molecular, Facultad de Ciencias Exactas Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina

    Pablo Bosch

  2. Regenerative Bioscience Center, The University of Georgia, Athens, GA

    Steven L. Stice

Authors
  1. Pablo Bosch
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  2. Steven L. Stice
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Editor information

Editors and Affiliations

  1. Stem Cells, Reprogenetics, West Orange, NJ

    Mohan C. Vemuri

  2. Invitrogen Corporation, Grand Island, NY

    Mohan C. Vemuri

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© 2007 Humana Press

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Bosch, P., Stice, S.L. (2007). Adenoviral Transduction of Mesenchymal Stem Cells. In: Vemuri, M.C. (eds) Stem Cell Assays. Methods in Molecular Biology™, vol 407. Humana Press. https://doi.org/10.1007/978-1-59745-536-7_18

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  • DOI: https://doi.org/10.1007/978-1-59745-536-7_18

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-744-0

  • Online ISBN: 978-1-59745-536-7

  • eBook Packages: Springer Protocols

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