Preparation of dendritic cells by in vitro cultures

Abstract

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 in fl ammatory-like DC using granulocyte-macrophages-colonystimulating factor (GM-CSF). These protocols allow for obtaining up to 2 × 10 8 CD11c high in fl ammatory-like DC and up to 5 × 10 6 equivalents of each CD8 + and CD8 - conventional DC and plasmacytoid DC. © Springer Science+Business Media, LLC 2013.