Protocol for mapping murine myeloid bone marrow progenitors and their differentiation into CD103+ cDC1s and CD301b+ cDC2s.

Ex vivo differentiation platforms have a long-standing history for studying dendritic cell (DC) ontogeny and function. Here, we present a protocol for differentiating bona fide DCs from murine bone marrow. We describe steps for bone marrow preparation, followed by ex vivo differentiation of DC subtypes resembling either lymphoid or mucosal tissue DC phenotypes. We then detail procedures for flow cytometric analysis and sorting strategies that facilitate phenotypic and functional studies of murine DC subtypes and/or their bone marrow progenitors. For complete details on the use and execution of this protocol, please refer to Amon et al.1.