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Updated: Jul 23, 2026
Phenotypic Analysis and Isolation of Murine Hematopoietic Stem Cells and Lineage-committed Progenitors
Published on: July 8, 2012
CD1a and CD1c cell sorting yields a homogeneous population of immature human Langerhans cells
Matthias Peiser1, Andreas Grützkau, Reinhard Wanner
1Department of Dermatology and Allergy, Charité, Humboldt-University of Berlin, D-10117 Berlin, Germany.
Insights
Researchers developed a new method to isolate human epidermal Langerhans cells (LCs) using CD1c selection. This protocol yields pure, viable immature dendritic cells (DCs) crucial for studying skin immunity and developing vaccines.
Area of Science:
- Immunology
- Dermatology
- Cell Biology
Background:
- Ex vivo generated Langerhans cells (LCs) have limitations in fully representing epidermal LCs for immunobiology studies.
- Developing effective methods to isolate pure, functional epidermal LCs is essential for research.
Purpose of the Study:
- To establish CD1-based magnetic-activated cell sorting (MACS) protocols for isolating human epidermal LCs.
- To characterize the phenotype, function, and viability of isolated LCs.
Main Methods:
- Utilized CD1c-based magnetic-activated cell sorting (MACS) for isolating human epidermal LCs.
- Assessed cell purity, viability, ultrastructural features, surface antigen expression, and cytokine profiles.
- Evaluated functional capacity using allogeneic mixed lymphocyte reactions and response to stimuli like LPS and IFN-omega.
Main Results:
- CD1c selection successfully isolated pure, viable HLA-DR(+)/CD1a(+) LCs with characteristic immature dendritic cell (DC) features.
- Demonstrated constitutive expression of CXCR4, CD40 ligand (CD40L), Fas, and Fas ligand (FasL) in normal LCs.
- Showed that isolated LCs maintain their immature state and viability in culture, with inducible inflammatory cytokine production.
Conclusions:
- The CD1c MACS protocol provides optimal human epidermal LCs for studying immature cell properties.
- These isolated LCs are suitable for developing immunotherapeutic vaccines.
- The study identified novel markers and mechanisms maintaining LC viability and immaturity.
Abstract:
There is increasing evidence that ex vivo generated Langerhans cells (LCs) cannot fully substitute for their physiological counterparts in normal epidermis when studying the immunobiology of this prototype of a tissue-residing immature dendritic cell (DC). Here, we present CD1-based magnetic-activated cell-sorting (MACS) protocols for the effective isolation of human epidermal LCs. CD1c selection yielded a homogeneous population of pure and viable HLA-DR(+)/CD1a(+) DCs, with the ultrastructural features, surface antigen expression and cytokine profile, characteristic of epidermis-resident immature LCs. The immature state and functional integrity were established by allogeneic mixed lymphocyte reactions showing a weak stimulatory capacity of freshly isolated cells and upregulation upon stimulation. Characterizing the cells in more detail, we could demonstrate for the first time that normal human LCs express CXCR4, CD40 ligand (CD40L), and Fas and Fas ligand (FasL). The observed constitutive transcription of TGF-beta suggests that the viability and immature state of epidermal LCs are maintained not only by the TGF-beta production from the microenvironment, but also in an autocrine or paracrine manner. LPS and IFN-omega stimulated the expression of the inflammatory cytokines TNF-alpha and IL-1beta, and there was secretion of IL-12p70 after CD40 ligation. Remarkably, the CD1-sorted LCs showed no loss of their Birbeck granules and CD1a expression upon culturing and no spontaneous phenotypic and functional maturation into potent antigen-presenting cells (APCs). We conclude that human epidermal LCs obtained by the CD1c cell-sorting protocol are optimal candidates with which to elucidate the properties and capabilities of immature cells and to develop immunotherapeutic vaccines.
