Related Experiment Video
Updated: Mar 21, 2026
Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture
Published on: April 8, 2016
Isolation of Human Skin Dendritic Cell Subsets
Merry Gunawan1, Laura Jardine1, Muzlifah Haniffa2
1Human DC Lab, Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
Insights
Human skin contains abundant dendritic cells (DCs) and macrophages, making skin explants a valuable source for studying these immune cells. Protocols are provided to isolate and identify skin DCs and macrophages using flow cytometry.
Area of Science:
- Immunology
- Dermatology
Background:
- Dendritic cells (DCs) are crucial leukocytes for antigen processing and presentation.
- DCs exist in various subsets, differing by location, phenotype, and function.
- Human skin harbors significantly more DCs than peripheral blood, with a more mature phenotype.
Purpose of the Study:
- To detail protocols for isolating dendritic cells (DCs) and resident macrophages from human skin.
- To provide a multiparameter flow cytometry gating strategy for identifying human skin DCs and distinguishing them from macrophages.
Main Methods:
- Isolation of dendritic cells (DCs) and macrophages from human skin explants.
- Utilization of multiparameter flow cytometry for cell identification and characterization.
Main Results:
- Skin explants offer an accessible source of mature dendritic cells (DCs) and macrophages.
- A reliable flow cytometry gating strategy enables accurate identification of skin DCs and differentiation from macrophages.
Conclusions:
- Human skin is a rich source of dendritic cells (DCs) and macrophages, ideal for immunological studies.
- Established protocols and flow cytometry methods facilitate the isolation and identification of these critical immune cells from skin.
Abstract:
Dendritic cells (DCs) are specialized leukocytes with antigen-processing and antigen-presenting functions. DCs can be divided into distinct subsets by anatomical location, phenotype and function. In human, the two most accessible tissues to study leukocytes are peripheral blood and skin. DCs are rare in human peripheral blood (<1 % of mononuclear cells) and have a less mature phenotype than their tissue counterparts (MacDonald et al., Blood. 100:4512-4520, 2002; Haniffa et al., Immunity 37:60-73, 2012). In contrast, the skin covering an average total surface area of 1.8 m(2) has approximately tenfold more DCs than the average 5 L of total blood volume (Wang et al., J Invest Dermatol 134:965-974, 2014). DCs migrate spontaneously from skin explants cultured ex vivo, which provide an easy method of cell isolation (Larsen et al., J Exp Med 172:1483-1493, 1990; Lenz et al., J Clin Invest 92:2587-2596, 1993; Nestle et al., J Immunol 151:6535-6545, 1993). These factors led to the extensive use of skin DCs as the "prototype" migratory DCs in human studies. In this chapter, we detail the protocols to isolate DCs and resident macrophages from human skin. We also provide a multiparameter flow cytometry gating strategy to identify human skin DCs and to distinguish them from macrophages.
