title: Isolation of dendritic cells.
abstract: This unit presents two methods for preparing dendritic cells (DCs), a highly specialized type of antigen-presenting cell (APC). The first method involves the isolation of DCs from mouse spleen, resulting in a cell population that is highly enriched in accessory cell and APC function. A support protocol for collagenase digestion of splenocyte suspensions is described to increase the yield of dendritic cells. The second method involves generating large numbers of DCs from mouse bone marrow progenitor cells. In that technique, bone marrow cells are cultured in the presence of granulocyte/macrophage colony-stimulating factor (GM-CSF) to yield 5-10 10(6) cells, 60% of which express DC surface markers (e.g., B-7-2/CD86). Additional techniques for isolating DCs from mouse spleens or other mouse tissues, as well as from human tissues, are also discussed.
authors.affiliation: Kyoto University, Kyoto, Japan.
authors.forename: Kayo
authors.initials: K
authors.lastname: Inaba
authors.affiliation: The Rockefeller University, New York, New York.
authors.forename: William J
authors.initials: WJ
authors.lastname: Swiggard
authors.affiliation: The Rockefeller University, New York, New York.
authors.forename: Ralph M
authors.initials: RM
authors.lastname: Steinman
authors.affiliation: University of Innsbruck, Innsbruck, Austria.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Romani
authors.affiliation: University of Erlangen, Erlangen, Germany.
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
mesh_terms: D000818:Animals; D051940:B7-2 Antigen; D001853:Bone Marrow / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases; D003470:Culture Media; D003713:Dendritic Cells / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D051379:Mice; D013154:Spleen / Q000276:immunology; D013234:Stem Cells / Q000276:immunology
chemical_list: D051940:B7-2 Antigen; D003470:Culture Media; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D017364:Collagenases