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Isolation and Characterization of Lymphocytes from Human Mucosal Biopsies

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Allergy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2020))

Abstract

The important role of the local mucosal environment in both the initiation and progression of allergic disease is well established. Analysis of tissue-resident lymphocyte subsets by flow cytometry requires isolation of viable cells from mucosal samples.

Here we describe an advanced method to dissociate lymphocytes from human mucosal (e.g., nasal, bronchial) biopsies. Single-cell suspensions are obtained through a combination of gentle mechanical disruption and incubation of tissue with proteolytic enzymes. This method fully utilizes limited clinical samples and is amenable to a variety of downstream applications for phenotypic, single-cell analysis of tissue lymphocytes or pooled lymphocyte subsets.

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Author information

Authors and Affiliations

  1. Asthma and Allergy, Randall Division of Cell and Molecular Biophysics, King’s College London, London, UK

    Nadine E. G. Upton

  2. MRC and Asthma UK Centre for Allergic Mechanisms of Asthma, King’s College London, Guy’s Campus, London, UK

    Nadine E. G. Upton

  3. Cell Motility and Cytoskeleton, Randall Division of Cell and Molecular Biophysics, King’s College London, London, UK

    Thomas S. Hayday

  4. Blizard Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK

    Louisa K. James

Authors
  1. Nadine E. G. Upton
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  2. Thomas S. Hayday
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  3. Louisa K. James
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Corresponding author

Correspondence to Louisa K. James .

Editor information

Editors and Affiliations

  1. Core Facilities, St. George’s University of London, London, UK

    Penny Lympany

  2. Department of Occupational and Environmental Medicine, Imperial College London, London, UK

    Meinir G. Jones

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© 2019 Springer Science+Business Media, LLC, part of Springer Nature

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Upton, N.E.G., Hayday, T.S., James, L.K. (2019). Isolation and Characterization of Lymphocytes from Human Mucosal Biopsies. In: Lympany, P., Jones, M. (eds) Allergy. Methods in Molecular Biology, vol 2020. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9591-2_12

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  • DOI: https://doi.org/10.1007/978-1-4939-9591-2_12

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9589-9

  • Online ISBN: 978-1-4939-9591-2

  • eBook Packages: Springer Protocols

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