Guidelines for DC preparation and flow cytometry analysis of mouse nonlymphoid tissues

Probst HC, Stoitzner P, Amon L, Backer RA, Brand A, Chen J, Clausen BE, Dieckmann S, Dudziak D, Heger L, Hodapp K, Hornsteiner F, Hovav AH, Jacobi L, Ji X, Kamenjarin N, Lahl K, Lahmar I, Lakus J, Lehmann C, Ortner D, Picard M, Roberti MP, Rossnagel L, Saba Y, Schalla C, Schlitzer A, Schraml BU, Schütze K, Seichter A, Seré K, Seretis A, Sopper S, Strandt H, Sykora MM, Theobald H, Tripp CH, Zitvogel L (2022)

Publication Type: Journal article

Publication year: 2022

Journal

DOI: 10.1002/eji.202249819

Abstract

This article is part of the Dendritic Cell Guidelines article series, which provides a collection of state-of-the-art protocols for the preparation, phenotype analysis by flow cytometry, generation, fluorescence microscopy and functional characterization of mouse and human dendritic cells (DC) from lymphoid organs and various nonlymphoid tissues. DC are sentinels of the immune system present in almost every mammalian organ. Since they represent a rare cell population, DC need to be extracted from organs with protocols that are specifically developed for each tissue. This article provides detailed protocols for the preparation of single-cell suspensions from various mouse nonlymphoid tissues, including skin, intestine, lung, kidney, mammary glands, oral mucosa and transplantable tumors. Furthermore, our guidelines include comprehensive protocols for multiplex flow cytometry analysis of DC subsets and feature top tricks for their proper discrimination from other myeloid cells. With this collection, we provide guidelines for in-depth analysis of DC subsets that will advance our understanding of their respective roles in healthy and diseased tissues. While all protocols were written by experienced scientists who routinely use them in their work, this article was also peer-reviewed by leading experts and approved by all coauthors, making it an essential resource for basic and clinical DC immunologists.

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Involved external institutions

Medizinische Universität Innsbruck Austria (AT) Institut Gustave-Roussy France (FR) Universitätsmedizin der Johannes Gutenberg-Universität Mainz Germany (DE) Rheinische Friedrich-Wilhelms-Universität Bonn Germany (DE) Leopold-Franzens-Universität Innsbruck / University of Innsbruck Austria (AT) Technical University of Denmark / Danmarks Tekniske Universitet (DTU) Denmark (DK) Hebrew University of Jerusalem Israel (IL) Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen Germany (DE) Klinikum der Universität München Germany (DE)

How to cite

APA:

Probst, H.C., Stoitzner, P., Amon, L., Backer, R.A., Brand, A., Chen, J.,... Zitvogel, L. (2022). Guidelines for DC preparation and flow cytometry analysis of mouse nonlymphoid tissues. European Journal of Immunology. https://doi.org/10.1002/eji.202249819

MLA:

Probst, Hans Christian, et al. "Guidelines for DC preparation and flow cytometry analysis of mouse nonlymphoid tissues." European Journal of Immunology (2022).

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