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Isolation of human skin dendritic cells by in vitro migration
C D Richters1, M J Hoekstra, J S du Pont
1Department of Cell Biology and Immunology, Vrije Universiteit, Amsterdam, The Netherlands.
Insights
Researchers developed a novel method to isolate antigen-presenting cells (APCs), specifically dendritic cells (DCs), from human skin without using enzymes. This technique leverages the natural migratory behavior of skin DCs for immune response studies.
Area of Science:
- Immunology
- Dermatology
- Cell Biology
Background:
- Human skin harbors diverse antigen-presenting cells (APCs), including Langerhans cells (LCs) in the epidermis and dendritic cells (DCs) in the dermis.
- Skin DCs are crucial for initiating immune responses by capturing antigens and migrating to lymph nodes for T-cell activation.
- Conventional methods for isolating skin DCs involve enzymatic digestion, which may alter cell surface marker expression.
Purpose of the Study:
- To develop and describe a novel method for obtaining dendritic cells (DCs) from human skin without enzymatic treatment.
- To characterize the isolated DCs and confirm their mature phenotype and functional potential.
Main Methods:
- Utilizing the intrinsic migratory capacity of skin DCs by culturing skin explants.
- Observing spontaneous migration of DCs out of the skin tissue into culture medium.
- Characterizing the migrated cells using established markers for dendritic cells and assessing their maturation state.
Main Results:
- A method was established to isolate DCs from human skin without enzymatic digestion.
- The spontaneously migrated cells exhibit characteristics of mature dendritic cells.
- The marker expression on these DCs is not influenced by enzymatic processing, providing a more accurate representation of their in vivo state.
Conclusions:
- This enzyme-free method provides a reliable way to obtain mature skin DCs.
- The technique preserves the natural phenotype of DCs, crucial for accurate immunological studies.
- This approach offers a valuable alternative for research involving skin-derived dendritic cells and immune responses.
Abstract:
In the human skin, various types of antigen-presenting cells (APC) are present. In the epidermis, they are identified ultrastructurally as Langerhans cells (LC) by the presence of Birbeck granules. LC are considered to belong to the family of dendritic cells (DC) that are important for the initiation of immune responses (1). In the dermis, macrophages and DC are present (2,3). The expression of CD1a molecules can be used to identify DC in the skin (4,5), because macrophages do not express this marker. In vivo, these skin DC are supposed to take up antigens penetrating in the skin. Thereafter, they migrate via the afferent lymphatics into the draining lymph nodes, where a T-cell response can be initiated (6,7). During migration, the DC mature into potent APC. Besides an increase in MHC class II expression, adhesion (8,9) and B7 co-stimulatory molecules (10) are up-regulated. Most research on skin DC has been carried out with cells isolated from enzyme digested skin (8-10). In this chapter, we describe a method to obtain DC from human skin without enzymes, by making use of their migratory capacities. The cells migrate "spontaneously" out of the skin during culture. Characterization of the cells shows that mature DC are obtained with a marker expression not influenced by enzymes.
