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In Vitro Methods for the Study of Glioblastoma Stem-Like Cell Radiosensitivity

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In Vitro Models for Stem Cell Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2269))

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Abstract

Ionizing radiation is a critical component of glioblastoma (GBM) therapy. Recent data have implicated glioblastoma stem-like cells (GSCs) as determinants of GBM development, maintenance, and treatment response. Understanding the response of GSCs to radiation should thus provide insight into the development of improved GBM treatment strategies. Towards this end, in vitro techniques for the analysis of GSC radiosensitivity are an essential starting point. One such method, the clonogenic survival assay has been adapted to assessing the intrinsic radiosensitivity of GSCs and is described here. As an alternative method, the limiting dilution assay is presented for defining the radiosensitivity of GSC lines that do not form colonies or only grow as neurospheres. In addition to these cellular strategies, we describe γH2AX foci analysis, which provides a surrogate marker for radiosensitivity at the molecular level. Taken together, the in vitro methods presented here provide tools for defining intrinsic radiosensitivity of GSCs and for testing agents that may enhance GBM radioresponse.

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Acknowledgments

JHM is supported by the NIH OxCam and Gates Cambridge Scholarships.

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Authors and Affiliations

  1. Radiation Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Joseph H. McAbee, Charlotte Degorre-Kerbaul & Philip J. Tofilon

  2. Department of Clinical Neurosciences, University of Cambridge, Cambridge, UK

    Joseph H. McAbee

  3. Wake Forest School of Medicine, Winston-Salem, NC, USA

    Joseph H. McAbee

Authors
  1. Joseph H. McAbee
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  2. Charlotte Degorre-Kerbaul
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  3. Philip J. Tofilon
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Corresponding author

Correspondence to Philip J. Tofilon .

Editor information

Editors and Affiliations

  1. Department of Visceral, Transplant, Thoracic and Vascular Surgery, Applied Molecular Hepatology Lab, University of Leipzig Medical Center, Leipzig, Germany

    Peggy Stock

  2. Department of Visceral, Transplant, Thoracic and Vascular Surgery, Applied Molecular Hepatology Lab, University of Leipzig Medical Center, Leipzig, Germany

    Bruno Christ

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McAbee, J.H., Degorre-Kerbaul, C., Tofilon, P.J. (2021). In Vitro Methods for the Study of Glioblastoma Stem-Like Cell Radiosensitivity. In: Stock, P., Christ, B. (eds) In Vitro Models for Stem Cell Therapy. Methods in Molecular Biology, vol 2269. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1225-5_3

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  • DOI: https://doi.org/10.1007/978-1-0716-1225-5_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1224-8

  • Online ISBN: 978-1-0716-1225-5

  • eBook Packages: Springer Protocols

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