Unraveling dimethylformamide-induced neutrophilic differentiation in HL-60 cells: A proteomic and functional comparison with dimethyl sulfoxide

by Othman Eldalal, Yasser Tabana, Dinesh Babu, Newton H. Tran, Steven Lockhart, Joshua Kranrod, John M.

Seubert, Lusine Tonoyan, Richard P. Fahlman, Arno G.

Siraki The differentiation of HL-60 cells into neutrophil-like cells is widely used to study neutrophil functions, yet no comprehensive proteomic analysis has been conducted on dimethylformamide (DMF)-induced differentiation. This study provides the first detailed proteomic characterization of DMF-differentiated (df)-HL-60 cells, demonstrating its distinct molecular and functional profiles compared to the well-established dimethyl sulfoxide (DMSO)-df-HL-60 cell model.

HL-60 cells were differentiated using 1.25% DMSO or 70 mM DMF for five days. Cell proliferation, granulocytic differentiation (CD11b expression), superoxide anion production, myeloperoxidase (MPO) protein expression and enzymatic activity, and neutrophil extracellular trap (NET) formation were evaluated.

Proteomic profiling was performed using LC-MS/MS, followed by gene ontology and pathway enrichment analysis to identify key molecular changes associated with differentiation. DMF-df-HL-60 cells maintained higher proliferation rates than DMSO-df-HL-60 cells.

Both agents successfully induced granulocytic differentiation, with DMSO producing greater CD11b expression.