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Isolation of Melanoma Cell Subpopulations Using Negative Selection

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Molecular Diagnostics for Melanoma

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1102))

Abstract

Melanomas are phenotypically and functionally heterogeneous tumors comprising of distinct subpopulations that drive disease progression and are responsible for resistance to therapy. Identification and characterization of such subpopulations are highly important to develop novel targeted therapies. However, this can be a challenging task as there is a lack of clearly defined markers to distinguish the melanoma subpopulations from a general tumor cell population. Also, there is a lack of optimal isolation methods and functional assays that can fully recapitulate their phenotype. Here we describe a method for isolating tumor cells from fresh human tumor tissue specimens using an antibody coupled magnetic bead sorting technique that is well established in our laboratory. Thus, melanoma cells are enriched by negative cell sorting and elimination of non-tumor cell population such as erythrocytes, leukocytes, and endothelial cells. Enriched unmodified tumor cells can be further used for phenotypic and functional characterization of melanoma subpopulations.

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Author information

Authors and Affiliations

  1. The Wistar Institute, Philadelphia, PA, USA

    Ana Slipicevic, Rajasekharan Somasundaram & Katrin Sproesser

  2. Department of Pathology, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway

    Ana Slipicevic

  3. Wistar Institute, Philadelphia, PA, USA

    Meenhard Herlyn

Authors
  1. Ana Slipicevic
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  2. Rajasekharan Somasundaram
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  3. Katrin Sproesser
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  4. Meenhard Herlyn
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Editor information

Editors and Affiliations

  1. Cancer Diagnosis Program, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA

    Magdalena Thurin

  2. Sidra Medical and Research Center,Qatar Foundation, Doha, Qatar

    Francesco M. Marincola

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© 2014 Springer Science+Business Media, New York

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Slipicevic, A., Somasundaram, R., Sproesser, K., Herlyn, M. (2014). Isolation of Melanoma Cell Subpopulations Using Negative Selection. In: Thurin, M., Marincola, F. (eds) Molecular Diagnostics for Melanoma. Methods in Molecular Biology, vol 1102. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-727-3_26

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  • DOI: https://doi.org/10.1007/978-1-62703-727-3_26

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-726-6

  • Online ISBN: 978-1-62703-727-3

  • eBook Packages: Springer Protocols

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