Isolation and Analysis of Mouse and Human Skin γδ T Cells - Cytometry Now

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Isolation and Analysis of Mouse and Human Skin γδ T Cells

Skin-resident and infiltrating ?δ T lymphocytes are components of the cutaneous immune system that provide the first line of defense against pathogens and the environment. Research that employs the isolation and culture of T cells from murine and human skin can help delineate the molecular and cellular mechanisms utilized by T lymphocytes in skin-specific immunity. However, obtaining high numbers of T cells from epithelial tissue without resorting to long-term culture or transformation can be difficult. Here, specific approaches are described for the isolation and culture of ?δ T lymphocytes from murine skin and human skin explant cultures. In addition, a protocol to assess the morphology and activation of epidermal ?δ T cells in situ using immunofluorescent microscopy is detailed. These techniques can be used to analyze resident and infiltrating ?δ T lymphocytes in the skin via flow cytometry, RNA-seq, or proteomics to further study inflammatory diseases, cancer, or autoimmunity. ? 2019 by John Wiley & Sons, Inc. Basic Protocol 1: Isolation, culture, and analysis of ?δ T cells from murine epidermis Basic Protocol 2: Examination of ?δ T cells in epidermal sheets to assess activation and morphology Basic Protocol 3: Preparation of human skin explant cultures for analysis of skin T cells Support Protocol 1: Counting live cells with hemocytometer Support Protocol 2: Preparing a Matrigel

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