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Lookup NU author(s): Dr Venetia BigleyORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2022 The Authors. European Journal of Immunology published by Wiley-VCH GmbH. 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.
Author(s): Lutz MB, Ali S, Audiger C, Autenrieth SE, Berod L, Bigley V, Cyran L, Dalod M, Dorrie J, Dudziak D, Florez-Grau G, Giusiano L, Godoy GJ, Heuer M, Krug AB, Lehmann CHK, Mayer CT, Naik SH, Scheu S, Schreibelt G, Segura E, Sere K, Sparwasser T, Tel J, Xu H, Zenke M
Publication type: Article
Publication status: Published
Journal: European Journal of Immunology
Year: 2023
Volume: 53
Issue: 11
Print publication date: 01/11/2023
Online publication date: 27/10/2022
Acceptance date: 10/08/2022
Date deposited: 22/11/2022
ISSN (print): 0014-2980
ISSN (electronic): 1521-4141
Publisher: John Wiley and Sons Inc.
URL: https://doi.org/10.1002/eji.202249816
DOI: 10.1002/eji.202249816
PubMed id: 36303448
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