RAW 264.7 cells (American Type Culture Collection [ATCC]) were cultured in DMEM (HyClone; GE Healthcare Life Science, Chicago, IL, USA) and THP-1 cells (ATCC) in RPMI 1640-GlutaMAX-I (Gibco; Life Technologies ltd, Inchinnan, UK); both media were supplemented with 10% (v/v) heat-inactivated FBS (FBSi; Invitrogen, Waltham, MA, USA) and 1% (v/v) antibiotic-antimycotic solution (AA; Invitrogen), at 37°C in 5% CO2.
Human CD14+ monocytes were isolated from blood from healthy donors and incubated in RPMI 1640 medium with 2 mM glutamax-I/glutamine (Gibco), 10% (v/v) FBSi, and 1% (v/v) AA. To obtain monocyte-derived pro-inflammatory macrophages (M1), primary monocytes were stimulated with 5 ng/ml GM-CSF (Sigma) for 7 days at 37°C in 5% CO2 as described previously [24 (link)].
Human CD14+ monocytes were isolated from blood from healthy donors and incubated in RPMI 1640 medium with 2 mM glutamax-I/glutamine (Gibco), 10% (v/v) FBSi, and 1% (v/v) AA. To obtain monocyte-derived pro-inflammatory macrophages (M1), primary monocytes were stimulated with 5 ng/ml GM-CSF (Sigma) for 7 days at 37°C in 5% CO2 as described previously [24 (link)].
