Labeling and tracking of immune cells in ex vivo human skin

Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled...

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Vydáno v:Nature protocols Ročník 16; číslo 2; s. 791 - 811
Hlavní autoři: Dijkgraaf, Feline E., Toebes, Mireille, Hoogenboezem, Mark, Mertz, Marjolijn, Vredevoogd, David W., Matos, Tiago R., Teunissen, Marcel B. M., Luiten, Rosalie M., Schumacher, Ton N.
Médium: Journal Article
Jazyk:angličtina
Vydáno: London Nature Publishing Group UK 01.02.2021
Nature Publishing Group
Témata:
631/1647/245/2186
631/1647/328/1978
631/1647/328/2057
631/1647/767/2202
631/250/1619/554/1834
Analytical Chemistry
Animals
Antibodies
Biological Techniques
Biomedical and Life Sciences
Biopsy
Biopsy - methods
CD4-Positive T-Lymphocytes - immunology
CD8 antigen
CD8-Positive T-Lymphocytes - immunology
Cell culture
Cell Culture Techniques - methods
Cell interaction
Cell research
Cell surface
Computational Biology/Bioinformatics
Health aspects
Humans
Identification and classification
Imaging
Immune system
Immunological memory
Labeling
Life Sciences
Lymphocytes
Lymphocytes T
Markers
Memory cells
Methods
Mice
Microarrays
Microscope and microscopy
Nanobodies
Nanotechnology
Organic Chemistry
Physiological aspects
Protocol
Reagents
Skin
Skin - cytology
Skin - immunology
Skin - pathology
Skin diseases
Staining and Labeling - methods
Surface markers
T cells
Tracking
Netherlands
ISSN:1754-2189, 1750-2799, 1750-2799
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Popis
Shrnutí:Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8 + T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8 + T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d. This protocol describes how to follow the behavior of immune cells by imaging ex vivo cultured human or mouse skin biopsies following labeling with antibody or nanobody reagents against specific cell surface markers.
Bibliografie:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:1754-2189
1750-2799
1750-2799
DOI:10.1038/s41596-020-00435-8