Isolation and Analysis of Mouse and Human Skin γδ T Cells.
Curr Protoc Immunol
; 127(1): e92, 2019 12.
Article
em En
| MEDLINE
| ID: mdl-31763791
ABSTRACT
Skin-resident and infiltrating γδ 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 γδ T lymphocytes from murine skin and human skin explant cultures. In addition, a protocol to assess the morphology and activation of epidermal γδ T cells in situ using immunofluorescent microscopy is detailed. These techniques can be used to analyze resident and infiltrating γδ 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 γδ T cells from murine epidermis Basic Protocol 2 Examination of γδ 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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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Pele
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Linfócitos T
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Separação Celular
Limite:
Animals
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Female
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Humans
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Male
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article