Isolated monocytes (from males) were plated onto tissue culture treated 6-wells at 2 × 106 cells/ml in RPMI-1640 media at 37°C 5%CO2 incubator. After two hours, media was aspirated to waste and adherent monocytes washed three times with DPBS and replaced with complete media composed of RPMI-1640, FBS (10% v/v), Penicillin/streptomycin (1% v/v), Glutamax (1X). To generate MD-Mϕ, M-CSF (25 ng/ml) was added to wells and cells differentiated for 5 days.
