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Identification and Immunophenotypic Characterization of Normal and Pathological Mast Cells

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Basophils and Mast Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2163))

Abstract

Mast cells (MCs) are secretory cells that are central players in human allergic disease and immune responses. With the exception of a few pathological situations, MCs are usually present at relatively low frequencies in most tissues. Since their first description, MCs in tissues were identified mostly using their morphological characteristics and their typical coloration when stained with aniline dyes. However, increasing availability of highly specific antibodies now permits the use of fluorescence-based flow cytometry as the method of choice for the quantification, characterization, and purification of cells in suspension. This technique allows for a rapid analysis of thousands of events and for the identification of cells present at frequencies as low as one event in 106 unwanted cells. This method also permits for simultaneous characterization of multiple antigens at a single cell level, which is ideal in order to study rare populations of cells like MCs. Here we describe the basis of flow cytometry-based immunophenotyping applied to the study of MC. The protocol focuses on the study of human MCs present in body fluids (mainly bone marrow) but can easily be adapted to studying MCs from other tissues and species.

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Authors and Affiliations

  1. Instituto de Estudios de Mastocitosis de Castilla La Mancha, Hospital Virgen del Valle, Toledo, Spain

    José Mário Morgado, Laura Sánchez-Muñoz & Luis María Escribano Mora

  2. Spanish Mastocytosis Network (REMA), Salamanca, Spain

    José Mário Morgado, Laura Sánchez-Muñoz, Cristina Teodósio & Luis María Escribano Mora

  3. Cytognos S.L. Biotechnology, Salamanca, Spain

    José Mário Morgado

  4. Centro de Investigación del Cáncer, Salamanca, Spain

    Cristina Teodósio & Luis María Escribano Mora

Authors
  1. José Mário Morgado
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  2. Laura Sánchez-Muñoz
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  3. Cristina Teodósio
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  4. Luis María Escribano Mora
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Editors and Affiliations

  1. Division of Experimental Allergology and Immunodermatology, Department of Human Medicine, School of Medicine and Health Sciences, Carl von Ossietzky University of Oldenburg, Oldenburg, Germany

    Bernhard F. Gibbs

  2. Institute for Parasitology, Justus Liebig University Giessen, Biomedizinisches Forschungszentrum Seltersberg (BFS), Giessen, Germany

    Franco H. Falcone

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Morgado, J.M., Sánchez-Muñoz, L., Teodósio, C., Mora, L.M.E. (2020). Identification and Immunophenotypic Characterization of Normal and Pathological Mast Cells. In: Gibbs, B.F., Falcone, F.H. (eds) Basophils and Mast Cells. Methods in Molecular Biology, vol 2163. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0696-4_27

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  • DOI: https://doi.org/10.1007/978-1-0716-0696-4_27

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0695-7

  • Online ISBN: 978-1-0716-0696-4

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