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Isolation of Cancer Stem Cells by Side Population Method

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Cancer Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1692))

Abstract

The Hoechst side population (SP) method is a flow cytometry technique used to obtain stem cells based on the dye efflux properties of the ATP-binding cassette (ABC) transporters. The SP cells are characterized by their capability to efflux the fluorescent DNA-binding dye Hoechst 33342 through their ABC transporters and are enriched in stem cells, which are endowed with a self-renewal capacity and multilineage differentiation potential and express the stemness genes including ABC multidrug transporters. The protocols outlined in this book chapter describe the isolation method of the SP cells from human lung carcinoma cell lines by using Hoechst 33342. In addition, we refer to the propagation method of SP cells by successive rounds of fluorescence-activated cell sorting analysis for SP cells. These approaches will be helpful for the establishment of novel in vitro and in vivo models using cancer stem cells, which may play a key role during carcinogenesis and/or tumor progression.

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Acknowledgments

This work was supported by Adaptable and Seamless Technology Transfer Program Through Target-driven R&G from the Japan Science and Technology Agency (AS2614150Q).

All the authors in this manuscript certify (1) that the paper has not been previously published or submitted elsewhere for publication; (2) that this study does not have financial or other relationships that might lead to a conflict of interest; (3) that all the authors have read and approved the manuscript, and also agreed with the authorship; (4) that Dr. Yasunori Okada is the corresponding author of this paper.

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Authors and Affiliations

  1. Department of Pathology, Keio University School of Medicine, Tokyo, Japan

    Masayuki Shimoda, Masahide Ota & Yasunori Okada

  2. Second Department of Internal Medicine and Respiratory Medicine, Nara Medical University, Nara, Japan

    Masahide Ota

  3. Department of Pathophysiology for Locomotive and Neoplastic Diseases, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo, 113-8421, Japan

    Yasunori Okada

Authors
  1. Masayuki Shimoda
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  2. Masahide Ota
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  3. Yasunori Okada
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Corresponding author

Correspondence to Yasunori Okada .

Editor information

Editors and Affiliations

  1. Department of Experimental Medicine, University of Campania “Luigi Vanvitelli”, Naples, Italy

    Gianpaolo Papaccio

  2. Department of Experimental Medicine, University of Campania “Luigi Vanvitelli”, Naples, Italy

    Vincenzo Desiderio

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Shimoda, M., Ota, M., Okada, Y. (2018). Isolation of Cancer Stem Cells by Side Population Method. In: Papaccio, G., Desiderio, V. (eds) Cancer Stem Cells. Methods in Molecular Biology, vol 1692. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7401-6_5

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  • DOI: https://doi.org/10.1007/978-1-4939-7401-6_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7400-9

  • Online ISBN: 978-1-4939-7401-6

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