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Using Retroviruses to Express Genes in Primary Megakaryocyte Lineage Cells

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Platelets and Megakaryocytes

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 273))

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Abstract

Megakaryocytes in adult human bone marrow are estimated to constitute approx 0.4% of the total marrow cells (1), and our experience suggests that fewer than 0.5% of low-density nucleated murine bone marrow cells express the megakaryocyte-lineage marker CD41 (integrin αIIb). Historically, the infrequent occurrence of megakaryocyte-lineage cells in bone marrow has been a significant obstacle to the procurement of primary megakaryocyte-lineage cells for biological studies. However, currently available conditions allow one to expand these cells in culture. In this chapter we describe protocols for using retroviruses to selectively infect early megakaryocyte-lineage cells and to infect mature megakaryocytes. The protocols allow one to study the effect of specific gene products on lineage development and biological functions of these primary cells. A basic understanding of retroviruses and the retrovirus life cycle is assumed (for review, see refs. 2,3).

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Author information

Authors and Affiliations

  1. Departments of Laboratory and Internal Medicine, University of California, San Francisco, CA

    Meenakshi Gaur & Andrew D. Leavitt

  2. Department of Genetics, Children’s Hospital, Harvard Medical School, Boston, MA

    George J. Murphy

  3. Department of Anatomy, Birmingham University Medical School, Birmingham, UK

    Jonathan Frampton

Authors
  1. Meenakshi Gaur
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  2. George J. Murphy
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  3. Jonathan Frampton
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  4. Andrew D. Leavitt
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Editor information

Editors and Affiliations

  1. School of Animal and Microbial Sciences, The University of Reading, Reading, UK

    Jonathan M. Gibbins

  2. Department of Physiology, University of Cambridge, Cambridge, UK

    Martyn P. Mahaut-Smith

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© 2004 Humana Press Inc.

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Gaur, M., Murphy, G.J., Frampton, J., Leavitt, A.D. (2004). Using Retroviruses to Express Genes in Primary Megakaryocyte Lineage Cells. In: Gibbins, J.M., Mahaut-Smith, M.P. (eds) Platelets and Megakaryocytes. Methods in Molecular Biology™, vol 273. Humana Press. https://doi.org/10.1385/1-59259-783-1:381

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  • DOI: https://doi.org/10.1385/1-59259-783-1:381

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-011-3

  • Online ISBN: 978-1-59259-783-3

  • eBook Packages: Springer Protocols

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