In Vitro Generation of Human Neutrophilic Myeloid-Derived Suppressor Cells

Myeloid-derived suppressor cells (MDSC) are a heterogeneous population of cells of myeloid origin. MDSC are functionally defined by their capacity to suppress T, NK, and B cell responses and henceforth altering the disease outcome in various pathological conditions. MDSC are further subdivided into three distinct subsets: monocytic (M-) MDSC, neutrophilic or polymorphonuclear (PMN-) MDSC, and early-stage (e-) MDSC. However, since surface markers utilized to define MDSC are expressed on other myeloid cells too, it is mandatory to functionally assess the suppressive activity for characterizing these cells. Here, we provide a protocol for generation of PMN-MDSC in vitro from freshly isolated human peripheral blood mononuclear cells. These MDSC can be used further to perform functional assays to determine their immunosuppressive potential or test their activities in various biological conditions, for instance in infection and cancer.