Glioblastoma induces the recruitment and differentiation of hybrid neutrophils from skull bone marrow

Tumor-associated neutrophils (TANs) are an under-characterized population in glioblastoma. We use scRNA-seq to explore phenotypic diversity among these cells. Peripheral blood neutrophils (PBNs) and TANs were isolated from GBM patients via FACS. Cells were stained with fluorophore-conjugated anti-human CD45 (HI30), CD16 (3G8), and CD66b (6/40c), and live CD45+CD66b+CD16hi neutrophils were sorted on a 4-laser 6-channel Sony SH800 cell sorter using ‘normal purity’ settings.