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Lookup NU author(s): James Fletcher, Professor Muzlifah Haniffa
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© The Author(s). Mononuclear phagocytes are present in skin and mucosa and represent one of the first lines of defense against invading pathogens, which they detect via an array of pathogen-binding receptors expressed on their surface. However, their extraction from tissue is difficult, and the isolation technique used has functional consequences on the cells obtained. Here, we compare mononuclear phagocytes isolated from human skin using either enzymatic digestion or spontaneous migration. Cells isolated via enzymatic digestion are in an immature state, and all subsets are easily defined. However, cells isolated by spontaneous migration are in a mature state, and CD141 cross-presenting DCs (cDC1) are more difficult to define. Different pathogen-binding receptors are susceptible to cleavage by blends of collagenase, demonstrating that great care must be taken in choosing the correct enzyme blend to digest tissue if carrying out pathogen-interaction assays. Finally, we have optimized mononuclear phagocyte culture conditions to enhance their survival after liberation from the tissue.
Author(s): Botting RA, Bertram KM, Baharlou H, Sandgren KJ, Fletcher J, Rhodes JW, Rana H, Plasto TM, Wang XM, Lim JJK, Barnouti L, Kohout MP, Papadopoulos T, Merten S, Olbourne N, Cunningham AL, Haniffa M, Harman AN
Publication type: Article
Publication status: Published
Journal: Journal of Leukocyte Biology
Year: 2017
Volume: 101
Issue: 6
Pages: 1393-1403
Print publication date: 01/06/2017
Online publication date: 07/03/2017
Acceptance date: 14/02/2017
Date deposited: 21/06/2017
ISSN (print): 0741-5400
ISSN (electronic): 1938-3673
Publisher: Federation of American Societies for Experimental Biology
URL: https://doi.org/10.1189/jlb.4A1116-496R
DOI: 10.1189/jlb.4A1116-496R
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