Guidelines for mouse and human DC generation

Lutz MB, Ali S, Audiger C, Autenrieth SE, Berod L, Bigley V, Cyran L, Dalod M, Dörrie J, Dudziak D, Florez-Grau G, Giusiano L, Godoy GJ, Heuer M, Krug AB, Lehmann C, Mayer CT, Naik SH, Scheu S, Schreibelt G, Segura E, Sere K, Sparwasser T, Tel J, Xu H, Zenke M (2022)

Publication Type: Journal article

Publication year: 2022

Journal

European Journal of Immunology Wiley-VCH Verlag

DOI: 10.1002/eji.202249816

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 non-lymphoid tissues. This article provides protocols with top ticks and pitfalls for preparation and successful generation of mouse and human DC from different cellular sources, such as murine BM and HoxB8 cells, as well as human CD34(+) cells from cord blood, BM, and peripheral blood or peripheral blood monocytes. We describe murine cDC1, cDC2, and pDC generation with Flt3L and the generation of BM-derived DC with GM-CSF. Protocols for human DC generation focus on CD34(+) cell culture on OP9 cell layers for cDC1, cDC2, cDC3, and pDC subset generation and DC generation from peripheral blood monocytes (MoDC). Additional protocols include enrichment of murine DC subsets, CRISPR/Cas9 editing, and clinical grade human DC generation. 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 co-authors, making it an essential resource for basic and clinical DC immunologists.

Authors with CRIS profile

Jan Dörrie Department of Dermatology Diana Dudziak Professur für die Biologie Dendritischer Zellen Christian Lehmann Department of Dermatology

Additional Organisation(s)

Interdisziplinäres Zentrum für Klinische Forschung (IZKF)

Related research project(s)

A087: Role of DC heterogeneity and natural antibodies for the disease outcome in cutaneous Leishmania infection July 1, 2020 - Nov. 30, 2023 A087: Role of DC heterogeneity and natural antibodies for the disease outcome in cutaneous Leishmania infection Dec. 1, 2020 - Nov. 30, 2023

Involved external institutions

Julius-Maximilians-Universität Würzburg DE Germany (DE) Heinrich-Heine-Universität Düsseldorf DE Germany (DE) Walter and Eliza Hall Institute of Medical Research (WEHI) AU Australia (AU) Deutsches Krebsforschungszentrum (DKFZ) DE Germany (DE) Johannes Gutenberg-Universität Mainz (JGU) DE Germany (DE) Newcastle University GB United Kingdom (GB) Aix-Marseille University / Aix-Marseille Université FR France (FR) Radboud University Nijmegen Medical Centre / Radboudumc of voluit Radboud Universitair Medisch Centrum (UMC) NL Netherlands (NL) Ludwig-Maximilians-Universität (LMU) DE Germany (DE) National Cancer Institute (NCI) US United States (USA) (US) PSL Research University / Université de recherche Paris Sciences et Lettres FR France (FR) Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen DE Germany (DE) Eindhoven University of Technology / Technische Universiteit Eindhoven (TU/e) NL Netherlands (NL)

How to cite

APA:

Lutz, M.B., Ali, S., Audiger, C., Autenrieth, S.E., Berod, L., Bigley, V.,... Zenke, M. (2022). Guidelines for mouse and human DC generation. European Journal of Immunology. https://doi.org/10.1002/eji.202249816

MLA:

Lutz, Manfred B., et al. "Guidelines for mouse and human DC generation." European Journal of Immunology (2022).

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