Study of myeloid cells endowed with suppressive activity is an active field of research which has particular importance in cancer, in view of the negative regulatory capacity of these cells to the host's immune response. The expansion of these cells, called myeloid‐derived suppressor cells (MDSCs), has been documented in many models of tumor‐bearing mice and in patients with tumors of various origin, and their presence is associated with disease progression and reduced survival. For this reason, monitoring this type of cell expansion is of clinical importance, and flow cytometry is the technique of choice for their identification. Over the years, it has been demonstrated that MDSCs comprise a group of immature myeloid cells belonging both to monocytic and granulocytic lineages, with several stages of differentiation; their occurrence depends on tumor‐derived soluble factors, which guide their expansion and determine their block of differentiation. Because of their heterogeneous composition, accurate phenotyping of these cells requires a multicolor approach, so that the expansion of all MDSC subsets can be appreciated.

 This review article focuses on identifying MDSCs and discusses problems associated with phenotyping circulating and tumor‐associated MDSCs in humans and in mouse models. © 2014 The Authors Cytometry Part B: Clinical Cytometry Published by Wiley Periodicals, Inc.