Evaluating EPA's Effects on Cell Viability

Cell viability was determined as described previously with slight revision [17 (link)]. Briefly, the differentiated HL60 cells were plated in 96-well plates at an initial density of 8000 cells/well in the presence of 100 ng/mL lipopolysaccharide (LPS, Millipore-Sigma, St. Louis, MO, USA) and EPA, in the form of free or liposomes, was supplemented to final concentrations of 40 and 80 µg/mL. The cells were further cultured in a humidified atmosphere containing 5% CO₂. After incubation of 20 h, 10 μL of the cell proliferation reagent CCK-8 (Promega, Madison, WI, USA) was added in every experimental well. 4 h later, the absorption at 490 nm was measured by a microplate reader (SynergyNeo, BioTek, Winooski, VT, USA). All experiments were performed in hex plicate.