Cited by 52Immunotherapy and Immune ResponsesImmune Response and InflammationImmune cells in cancer

Phenotypic and functional consequences of different isolation protocols on skin mononuclear phagocytes

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Botting, Rachel A, Bertram, Kirstie M, Baharlou, Heeva, Sandgren, Kerrie J, Fletcher, James, Rhodes, Jake W, Rana, Hafsa, Plasto, Toby M +10 more·2017·Journal of Leukocyte Biology

Abstract

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.

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  1. [24]Carpentier S., Vu Manh T.P., Chelbi R., Henri S., Malissen B., Haniffa M., Ginhoux F., Dalod M. (2016) Comparative genomics analysis of mononuclear phagocyte subsets confirms homology between lymphoid tissue-resident and dermal XCR1+ DCs in mouse and human and distinguishes them from Langerhans cells. J. Immunol. Methods 432, 35– 49. 26966045 PMC4859332

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Authors
Botting, R. A., Bertram, K. M., Baharlou, H., Sandgren, K. J., Fletcher, J., Rhodes, J. W., Rana, H., Plasto, T. M., Wang, X. M., Lim, J. J. K., Barnouti, L., Kohout, M. P., Papadopoulos, T., Merten, S., Olbourne, N., Cunningham, A. L., Haniffa, M., & Harman, A. N.
Affiliations
  • The University of Sydney (AU)
  • Newcastle University (GB)
  • Westmead Institute for Medical Research (AU)
  • Parramatta , New South Wales , Australia
  • Australia Plastic Surgery , Sydney, New South Wales , Australia
  • Cosmetic Culture , Pyrmont, New South Wales , Australia
  • Pure Aesthetics Plastic Surgery , Sydney, New South Wales , Australia
  • Palm Beach , New South Wales , Australia
  • Royal Victoria Infirmary (GB)
Corresponding author
Harman, Andrew N
Venue
Journal of Leukocyte Biology
Published
2017-03-07
Volume
101
Issue
6
Pages
1393-1403
ISSN
  • 0741-5400 (ISSN-L)
  • 1938-3673
OpenAlex
W2592533921
Cited by
52
References
36
Citations by year
2026: 5 · 2025: 3 · 2024: 2 · 2023: 5 · 2022: 9 · 2021: 11 · 2020: 8 · 2019: 7 · 2018: 1 · 2017: 1
FWCI
2.10
Type
journal article
Language
en
Open access
Locations
  • Journal of Leukocyte Biology (publishedVersion, cc-by-nc)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Newcastle University ePrints (Newcastle Univesity) (cc-by)
Funders
National Health and Medical Research Council (WT107931/Z/15/Z)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
  • Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
  • Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
Keywords
monocyte isolation, enzymatic digestion, collagenase, spontaneous migration, cDC1, thrombomodulin, culture conditions
MeSH terms
Cell Movement, Cell Separation, Enzymes, Humans, Monocytes, Phagocytes, Phenotype, Skin

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