Generation of Large Numbers of Pro-DCs and Pre-DCs In Vitro

The CD8+ , CD8− , and plasmacytoid dendritic cell (DC) subtypes develop from progenitors that express surface fms -like tyrosine kinase 3 (Flt3). Recently, two developmentally sequential progenitors have been identified that give rise to these subtypes. This includes a transition from an earlier CD11c− MHC-II− “pro-DC,” which divides and differentiates to give rise to CD11c+ MHC-II− “pre-DC,” en route to generating the three CD11c+ MHC-II+ DC subtypes – plasmacytoid DCs, CD8+ DCs, and CD8− DCs. In this chapter, we describe the very simple method of generating large numbers of in vitro-derived pro-DCs and pre-DCs. As these precursors are largely DC-restricted, they can be used to either reconstitute a mouse with DCs of desired background, to study the developmental steps in vitro or in vivo, among other purposes.