Protocol for the differentiation of HL-60 cells into a neutrophil-like state

The human myeloid HL-60 cell line serves as a substitute model in neutrophil research. Here, we present a protocol for HL-60 neutrophil differentiation with combined DMSO/all-trans retinoic acid (ATRA) treatment for 5 days. We describe steps for treating and incubating HL-60 cells. We then detail procedures for harvesting and staining of differentiated HL-60 cells for validation of neutrophil maturation. For complete details on the use and execution of this protocol, please refer to Hornstein et al.¹^,².