title: Generation and Identification of GM-CSF Derived Alveolar-like Macrophages and Dendritic Cells From Mouse Bone Marrow.
abstract: Macrophages and dendritic cells (DCs) are innate immune cells found in tissues and lymphoid organs that play a key role in the defense against pathogens. However, they are difficult to isolate in sufficient numbers to study them in detail, therefore, in vitro models have been developed. In vitro cultures of bone marrow-derived macrophages and dendritic cells are well-established and valuable methods for immunological studies. Here, a method for culturing and identifying both DCs and macrophages from a single culture of primary mouse bone marrow cells using the cytokine granulocyte macrophage colony-stimulating factor (GM-CSF) is described. This protocol is based on the established procedure first developed by Lutz et al. in 1999 for bone marrow-derived DCs. The culture is heterogeneous, and MHCII and fluoresceinated hyaluronan (FL-HA) are used to distinguish macrophages from immature and mature DCs. These GM-CSF derived macrophages provide a convenient source of in vitro derived macrophages that closely resemble alveolar macrophages in both phenotype and function.
authors.affiliation: Microbiology and Immunology, University of British Columbia.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Microbiology and Immunology, University of British Columbia.
authors.forename: Arif A
authors.initials: AA
authors.lastname: Arif
authors.affiliation: Microbiology and Immunology, University of British Columbia.
authors.forename: Grace F T
authors.initials: GF
authors.lastname: Poon
authors.affiliation: Microbiology and Immunology, University of British Columbia.
authors.forename: Blair
authors.initials: B
authors.lastname: Hardman
authors.affiliation: Microbiology and Immunology, University of British Columbia.
authors.forename: Manisha
authors.initials: M
authors.lastname: Dosanjh
authors.affiliation: Microbiology and Immunology, University of British Columbia; pauline@mail.ubc.ca.
authors.forename: Pauline
authors.initials: P
authors.lastname: Johnson
mesh_terms: D000818:Animals; D001853:Bone Marrow; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002478:Cells, Cultured; D003713:Dendritic Cells; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D016173:Macrophage Colony-Stimulating Factor; D008264:Macrophages; D051379:Mice
chemical_list: D016173:Macrophage Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor