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Isolation of Live Immune Cells from the Tumor Microenvironment by FACS

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Cancer Immunotherapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2748))

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Abstract

Isolation of live cells from the tumor microenvironment (TME) has represented a challenge, particularly from metastatic nodules that need to be identified within the entire metastatic tissue. Cherry-niche, an in vivo labelling technique, allows the isolation of all the different cell populations in the TME without needing to visually locate the metastatic cancer cell colonies. Therefore, neighboring TME cells can be isolated even from the early stages of cancer cell seeding and colonization in the metastatic tissue. Here, we show how to use Cherry-niche to identify and isolate neutrophils from the lung metastatic niche. We also provide examples of downstream analyses to characterize freshly isolated neutrophils ex vivo, such as Giemsa staining, reactive oxygen species (ROS) detection, and phagocytosis assays. Similar strategies can be used to isolate other immune and non-immune cells from the metastatic TME.

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Author notes

  1. Aikaterini Kafka and Christos Ermogenous contributed equally to this work.

Authors and Affiliations

  1. Centre for Tumour Microenvironment, Barts Cancer Institute, Queen Mary University London, John Vane Science Centre, London, UK

    Aikaterini Kafka, Christos Ermogenous & Luigi Ombrato

Authors
  1. Aikaterini Kafka
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  2. Christos Ermogenous
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  3. Luigi Ombrato
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Corresponding author

Correspondence to Luigi Ombrato .

Editor information

Editors and Affiliations

  1. Synthetic and Systems Biology for Biomedicine, Isituto Italiano di Tecnologia-IIT, Napoli, Napoli, Italy

    Velia Siciliano

  2. Department of Chemical Engineering, Imperial College, London, UK

    Francesca Ceroni

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Kafka, A., Ermogenous, C., Ombrato, L. (2024). Isolation of Live Immune Cells from the Tumor Microenvironment by FACS. In: Siciliano, V., Ceroni, F. (eds) Cancer Immunotherapy. Methods in Molecular Biology, vol 2748. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3593-3_1

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  • DOI: https://doi.org/10.1007/978-1-0716-3593-3_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3592-6

  • Online ISBN: 978-1-0716-3593-3

  • eBook Packages: Springer Protocols

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