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Flow Cytometry Enumeration of Apoptotic Cancer Cells by Apoptotic Rate

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Apoptosis and Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1219))

Abstract

Most authors currently quantify the frequency of apoptotic cells in a given phenotypically defined population after calculating the apoptotic index (AI), i.e., the percentage of apoptotic cells displaying a specific linage antigen (LAg) within a population of cells that remain unfragmented and retain the expression of the LAg. However, this approach has two major limitations. Firstly, apoptotic cells fragment into apoptotic bodies that later disintegrate. Secondly, apoptotic cells frequently lose, partially or even completely, the cell surface expression of the LAg used for the identification of specific cell subsets. This chapter describes a flow cytometry method to calculate the apoptotic rate (AR) that takes into account both cell fragmentation and loss of lineage antigen expression on measurement of apoptosis using flow cytometry ratiometric cell enumeration that emerges as a more accurate method of measurement of the occurrence of apoptosis in normal and tumoral cell cultures.

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Author information

Authors and Affiliations

  1. CNB-CSIC R&D Associated Unit, Department of Medicine, University of Alcalá, Carretera Madrid-Barcelona, Km 33.600, 28871, Alcalá de Henares, Madrid, Spain

    David Diaz B.Sc., Alfredo Prieto B.Sc., Ph.D., Eduardo Reyes B.Sc., Ph.D., Hugo Barcenilla B.Sc., Jorge Monserrat B.Sc., Ph.D. & Melchor Alvarez-Mon M.D., Ph.D.

  2. Research Unit, Industrial Farmacéutica Cantabria, Madrid, Spain

    Eduardo Reyes B.Sc., Ph.D.

  3. Immune System Diseases and Oncology Service, University Hospital “Príncipe de Asturias”, Alcalá de Henares, Madrid, Spain

    Melchor Alvarez-Mon M.D., Ph.D.

Authors
  1. David Diaz B.Sc.
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  2. Alfredo Prieto B.Sc., Ph.D.
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  3. Eduardo Reyes B.Sc., Ph.D.
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  4. Hugo Barcenilla B.Sc.
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  5. Jorge Monserrat B.Sc., Ph.D.
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  6. Melchor Alvarez-Mon M.D., Ph.D.
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Corresponding author

Correspondence to David Diaz B.Sc. .

Editor information

Editors and Affiliations

  1. Department of Obstetrics, Gynecology, and Reproductive Sciences,, Yale University, New Haven, Connecticut, USA

    Gil Mor

  2. Department of Obstetrics, Gynecology, and Reproductive Sciences,, Yale University, New Haven, Connecticut, USA

    Ayesha B. Alvero

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Diaz, D., Prieto, A., Reyes, E., Barcenilla, H., Monserrat, J., Alvarez-Mon, M. (2015). Flow Cytometry Enumeration of Apoptotic Cancer Cells by Apoptotic Rate. In: Mor, G., Alvero, A. (eds) Apoptosis and Cancer. Methods in Molecular Biology, vol 1219. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1661-0_2

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  • DOI: https://doi.org/10.1007/978-1-4939-1661-0_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1660-3

  • Online ISBN: 978-1-4939-1661-0

  • eBook Packages: Springer Protocols

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