Isolation and characterization of migratory human skin dendritic cells.
A method is described to isolate and characterize human skin dendritic cells (DC). This method is based on the migratory capacities of these cells. The cells migrated 'spontaneously' out of split-skin explants into the medium during a 24-h culture period and contained up to 75% CD1a+ cells. After removal of co-migrated T cells and macrophages, the highly enriched (> 95% CD1a+) DC showed potent allo-antigen-presenting capacities. About 25% of the CD1a+ cells were also positive for the dermal DC marker CD1b, whereas only 15-20% of the cells contained Birbeck granules, the characteristic cell organelle of the epidermal Langerhans cell. Before culture, CD1a+ DC were observed on cryostat sections not only in the epidermis but also in the dermis. After culture, the number of CD1a+ cells in both epidermis and dermis had decreased. Not all the cells had migrated during the culture period; some CD1a+ cells could still be detected in the epidermis and dermis after culture. Thus, using this method, potent allo-stimulating CD1a+ cells, migrating from both epidermis and dermis, can be obtained without the use of enzymes.
Richters
C D
CD
Department of Cell Biology, Faculty of Medicine, Vrije Universiteit, Amsterdam, The Netherlands.
Hoekstra
M J
MJ
van Baare
J
J
Du Pont
J S
JS
Hoefsmit
E C
EC
Kamperdijk
E W
EW
D002465:Cell Movement / Q000276:immunology*; D002469:Cell Separation / Q000379:methods; D046508:Culture Techniques / Q000379:methods; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology* / Q000648:ultrastructure; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D007801:Langerhans Cells / Q000166:cytology / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D012867:Skin / Q000166:cytology* / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology