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Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture
Published on: April 8, 2016
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Isolation and Analysis of Mouse and Human Skin γδ T Cells
Shannon Gargas1, Savannah Bshara-Corson1, Michelle Cruz1
1Department of Biological Sciences, California State University, San Marcos, California.
Current Protocols in Immunology
|November 26, 2019
Summary
This study details methods for isolating and culturing gamma delta T lymphocytes from mouse and human skin. These protocols enable the analysis of these immune cells in skin immunity and disease research.
Area of Science:
- Immunology
- Dermatology
- Cell Biology
Background:
- Gamma delta (γδ) T lymphocytes are crucial for skin immunity, acting as a first defense against pathogens.
- Isolating sufficient γδ T cells from skin for research is challenging without long-term culture or transformation.
Purpose of the Study:
- To describe protocols for isolating and culturing γδ T lymphocytes from murine and human skin.
- To detail methods for assessing γδ T cell morphology and activation in situ.
- To provide techniques for analyzing skin-resident and infiltrating γδ T cells.
Main Methods:
- Isolation and culture of γδ T cells from murine epidermis.
- In situ examination of epidermal γδ T cells using immunofluorescent microscopy.
- Preparation of human skin explant cultures for T cell analysis.
Main Results:
- Established specific approaches for obtaining and culturing γδ T lymphocytes from skin.
- Developed a protocol for assessing epidermal γδ T cell morphology and activation.
- Provided methods applicable to flow cytometry, RNA-seq, and proteomics.
Conclusions:
- The described techniques facilitate the study of γδ T lymphocytes in skin immunity.
- These methods support research into inflammatory diseases, cancer, and autoimmunity involving skin T cells.
- The protocols enable detailed analysis of γδ T cells in various skin conditions.
