Preparation of Dendritic Cells by In Vitro Cultures

In vitro cultures of bone marrow-derived precursors are a convenient method for generating dendritic cells (DC). This method additionally overcomes the problem of low availability of certain DC types, DC heterogeneity, and laborious procedures encountered using ex vivo isolation protocols. Here we describe two standard protocols for in vitro differentiation of steady-state DC equivalents with Fms-like tyrosine kinase 3 ligand (Flt3L) and inflammatory-like DC using granulocyte-macrophages-colony-stimulating factor (GM-CSF). These protocols allow for obtaining up to 2 � 108 CD11chigh inflammatory-like DC and up to 5 � 106 equivalents of each CD8+ and CD8− conventional DC and plasmacytoid DC.