In Vitro Generation of Murine Myeloid-Derived Suppressor Cells, Analysis of Markers, Developmental Commitment, and Function

Myeloid-derived suppressor cells (MDSC) appear at relatively low frequencies in diseased organs such as tumors or infection sites, but accumulate systemically in the spleen. So far MDSC have been reported in humans and experimental animals such as mice, rats, and nonhuman primates. Therefore, methods to generate MDSC in large amounts in vitro can serve as an additional tool to study their biology. Here, we describe in detail the generation of murine MDSC with GM-CSF from bone marrow (BM). Both subsets of granulocytic (G-MDSC) and monocytic MDSC (M-MDSC) are generated by this cytokine. We provide panels of phenotypic markers to distinguish them from non-suppressive cells and define developmental stages of monocytes developing into M-MDSC by two subsequent steps in vitro.