P-FPMI-111 - P-FPMI-111
Human monocytic cells, THP-1, were obtained from American type culture collection, ATCC (TIB-202) and were grown in suspension in RPMI-1640 media (Gibco, Invitrogen Life technologies, Burlington, ON), supplemented with 10% (v/v) heat inactivated fetal bovine serum (FBS), 2 mM L-glutamine and 1 mM sodium pyruvate (all from Invitrogen Life Technologies). Cultures were maintained at 37 degree Celsius in a humidified 5% (v/v) CO2 incubator up to a maximum of six passages. THP-1 cells at a density of 1 x 106 cells /ml were treated with 0.3 ug/ml phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich Canada, Oakville ON) for 24 hrs, inducing plastic-adherent (primed) cells that were further rested in complete RPMI-1640 medium for an additional 24 hrs prior to stimulations with various treatments. All experiments using human THP-1 (primed) cells were done for at least three biological replicates with two technical replicates each.