Flow cytometry analysis of the subpopulations of mouse keratinocytes and skin immune cells.
Open accessAbstract
Skin is our body's outermost physical barrier and an immunological interface enriched with various immune and non-immune cells. However, efficient generation of single-cell suspensions for flow cytometry analysis can be challenging. Here, we provide protocols to obtain epidermal and whole skin cell suspensions as well as gating strategies to identify mouse keratinocytes and skin immune cell subsets via flow cytometry. For complete details on the use and execution of this protocol, please refer to Sakamoto et al. (2021).
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IL-1Ra induction may differ between murine and human skin, and intraspecies variation in the IL-1 regulation of innate immune response may exist.
“IL-1Ra induction may differ between the murine and human in the skin, and intraspecies variation in the IL-1 regulation of innate immune response may exist [46]”
From Single-cell transcriptomics unveils skin cell specific antifungal immune responses and IL-1Ra- IL-1R immune evasion strategies of emerging fungal pathogen Candida auris (2024) · cites this paper as [46]
The Expected outcomes section describes flow-cytometry gating for mouse skin immune cells, and the Limitations section discusses differences between mouse strains in NK1.1 expression. Neither addresses IL-1Ra induction in murine versus human skin or intraspecies variation in IL-1 regulation. Searches for IL-1Ra and interleukin-1 receptor antagonist found no relevant passage in our copy; page numbers were unavailable. (LLM, conf=0.76)
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References 11 · Cited by 24
Research works printed but not linked: 9
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- Azzi L. El-Alfy M. Martel C. Labrie F. Gender differences in mouse skin morphology and specific effects of sex steroids and dehydroepiandrosterone J. Invest. Dermatol. 124 2005 22 27 15654949
- Carlyle J.R. Mesci A. Ljutic B. Belanger S. Tai L.H. Rousselle E. Troke A.D. Proteau M.F. Makrigiannis A.P. Molecular and genetic basis for strain-dependent NK1.1 alloreactivity of mouse NK cells J. Immunol. 176 2006 7511 7524 16751398
- Kashem S.W. Kaplan D.H. Isolation of murine skin resident and migratory dendritic cells via enzymatic digestion Curr. Protoc. Immunol. 121 2018 e45 30040218 PMC6060630
- Ko Y.C. Chien H.F. Jiang-Shieh Y.F. Chang C.Y. Pai M.H. Huang J.P. Chen H.M. Wu C.H. Endothelial CD200 is heterogeneously distributed, regulated and involved in immune cell-endothelium interactions J. Anat. 214 2009 183 195 19166481 PMC2667927
- Müller-Röver S. Handjiski B. van der Veen C. Eichmüller S. Foitzik K. McKay I.A. Stenn K.S. Paus R. A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages J. Invest. Dermatol. 117 2001 3 15 11442744
- Nagao K. Ginhoux F. Leitner W.W. Motegi S. Bennett C.L. Clausen B.E. Merad M. Udey M.C. Murine epidermal Langerhans cells and Langerin-expressing dermal dendritic cells are unrelated and exhibit distinct functions Proc. Natl. Acad. Sci. U S A 106 2009 3312 3317 19218433 PMC2651335
- Sakamoto K. Jin S.P. Goel S. Jo J.H. Voisin B. Kim D. Nadella V. Liang H. Kobayashi T. Huang X. Disruption of the endopeptidase ADAM10-Notch signaling axis leads to skin dysbiosis and innate lymphoid cell-mediated hair follicle destruction Immunity 54 2021 2321 2337.e10 34582748 PMC8516731
- Schneider M.R. Schmidt-Ullrich R. Paus R. The hair follicle as a dynamic miniorgan Curr. Biol. 19 2009 R132 R142 19211055
- Authors
- Sakamoto, K., Goel, S., Funakoshi, A., Honda, T., & Nagao, K.
- Affiliations
- National Institutes of Health (US)
- Hamamatsu University (JP)
- Corresponding author
- Sakamoto, Keiko
- Nagao, Keisuke
- Venue
- STAR Protocols
- Publisher
- Elsevier BV
- Published
- 2022-03
- Volume
- 3
- Issue
- 1
- Pages
- 101052
- ISSN
- 2666-1667 (ISSN-L)
- PMC
- PMC8689364
- OpenAlex
- W4200221183
- Cited by
- 24
- References
- 11
- Citations by year
- 2026: 6 · 2025: 7 · 2024: 6 · 2023: 4 · 2022: 1
- FWCI
- 2.44
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- STAR Protocols (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Institute of Arthritis and Musculoskeletal and Skin Diseases
- Topics
- Health Sciences › Medicine › Dermatology › Dermatology and Skin Diseases
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Skin and Cellular Biology Research
- Keywords
- keratinocytes, skin immune cells, mouse skin, epidermis, cell suspension preparation, gating strategy, single-cell analysis, skin barrier
- MeSH terms
- Animals, Flow Cytometry, Skin, Suspensions, Keratinocytes, Mice
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