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Updated: Aug 8, 2025
Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture
Published on: April 8, 2016
Isolation of Lymphocytes from Human Skin and Murine Tissues: A Rapid and Epitope-Preserving Approach
Alexandra Polakova1, Christoph Hudemann1, Felix Wiemers2
1Department of Dermatology and Allergology, Philipps-Universität Marburg, Marburg, Germany.
Insights
Researchers developed a fast protocol to isolate immune cells from skin biopsies. This method enables detailed analysis of T cells and other lymphocytes for skin disease research and therapeutic target identification.
Area of Science:
- Immunology
- Dermatology
- Cell Biology
Background:
- Tissue-resident immune cells are crucial for skin health and disease.
- Characterizing skin-derived immune cells is challenging due to sample access and complex protocols.
- Blood leukocytes are often used as surrogates but may not represent local skin immunity.
Purpose of the Study:
- To establish a rapid protocol for isolating viable immune cells from small human skin biopsies.
- To enable comprehensive phenotyping and functional studies of T cells and other lymphocytes.
- To provide a method applicable to both human and murine skin and mucosa.
Main Methods:
- Optimization of enzymatic digestion using type IV collagenase and DNase I.
- Isolation of immune cells from 4-mm skin biopsies.
- Validation of cell viability and marker preservation for multicolor flow cytometry.
Main Results:
- The optimized protocol significantly increases cellular yield from skin biopsies.
- Enzyme combination preserves leukocyte markers essential for flow cytometry analysis.
- The protocol is effective for both human and murine skin and mucosal samples.
Conclusions:
- This rapid protocol facilitates the comprehensive analysis of skin-resident immune cells.
- It supports disease surveillance and the identification of therapeutic targets in skin conditions.
- Enables deeper characterization of lymphocyte subpopulations in skin immunity.
Abstract:
Tissue-resident immune cells have been shown to play an important role in skin health and disease. However, owing to limited access to human skin samples and time-consuming, technically demanding protocols, the characterization of tissue-derived cells remains challenging. For this reason, blood-derived leukocytes are frequently used as a surrogate specimen, although they do not necessarily reflect local immune responses in the skin. Therefore, we aimed to establish a rapid protocol to isolate a sufficient number of viable immune cells from 4-mm skin biopsies that can be directly used for a deeper characterization such as comprehensive phenotyping and functional studies of T cells. In this optimized protocol, only two enzymes, type IV collagenase and DNase I, were used to achieve both the highest possible cellular yield and marker preservation of leukocytes stained for multicolor flow cytometry. We further report that the optimized protocol may be used in the same manner for murine skin and mucosa. In summary, this study allows a rapid acquisition of lymphocytes from human or mouse skin suitable for comprehensive analysis of lymphocyte subpopulations, for disease surveillance, and for identification of potential therapeutic targets or other downstream applications.
