title: The isolation and identification of murine dendritic cell populations from lymphoid tissues and their production in culture.
abstract: Dendritic cells (DC) are widely regarded as the most potent cellular inducers of the adaptive immune response; so, immunotherapy through DC manipulation is a promising option in the future fight against many human ailments. We have developed a method of isolating DC from the mouse that involves efficient extraction from tissues, followed by the selection of the lightest density cells, then depletion of non-DC through a cocktail of monoclonal antibodies and anti-immunoglobulin magnetic beads. Finally, purification and segregation into DC subtypes is achieved by immunofluorescent labeling and sorting. This has demonstrated a network of DC populations differing in surface phenotype and function. We can now produce larger numbers of many of these DC subpopulations from their precursors using bone marrow cultures supplemented with fms-like tyrosine kinase 3 ligand (Flt3L). The culture-generated DC can be aligned with the populations directly isolated from tissues. Combining the in vivo and in vitro systems will make study of murine DC and their alignment to their human counterparts an easier break process.
authors: The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.
authors: David
authors: D
authors: Vremec
authors: Ken
authors: K
authors: Shortman
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000378:metabolism; D001853:Bone Marrow; D018929:Cell Culture Techniques / Q000379:methods*; D019070:Cell Lineage; D002469:Cell Separation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D018189:Immunomagnetic Separation; D008221:Lymphoid Tissue / Q000166:cytology*; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D008565:Membrane Proteins; C084881:flt3 ligand protein