Work

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

Year: 2025

Type: article

Abstract: 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 • Protocol to derive neutrophil-like cells from the human myeloid HL-60 cell line • Staining steps with PI or May-Gruenwald-Giemsa for validation of neutrophil maturation • Instructions for analysis of cell surface markers of differentiation and proliferation • Methods for ROS and phagocytosis measurement for evaluation of neutrophil functionality Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104135

Authors: Tamara Hornstein, Klaus Unfried

Institution: Leibniz Institute of Environmental Medicine

Language: English

Topic: Retinoids in leukemia and cellular processes

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550