Dmem hepes medium

Bone marrow cells were obtained by flushing the femurs from mice with Dulbecco's modified Eagle's medium (DMEM)‐HEPES medium (Gibco). Cells were collected in 50‐mL tubes and centrifuged for 10 minutes (100 g). The supernatant was removed, and cells were suspended in DMEM (20% FBS; 20% L929 supernatant). Then, 1 × 10⁶ cells were cultured in 6‐well plate at 37°C and 5% CO2 for 7 days (BMDMs). Macrophage polarization was obtained by removing the culture medium and culturing cells for an additional 48 hours in DMEM supplemented with 10% FBS and 100 ng/mL LPS (Sigma) plus 20 ng/mL IFN‐γ (PeproTech) (for M1 polarization) or 20 ng/mL IL‐4 (PeproTech) (for M2 polarization).

Bone marrow cells were obtained by flushing the femurs from mice with Dulbecco’s modified Eagle’s medium (DMEM)-HEPES medium (Gibco). Cells were collected in 50 ml tubes and centrifuged for 10 min (100 × g). The supernatant was removed, and cells were suspended in DMEM (10% FCS; 20% L929 supernatant). Then, 1 × 10⁶ cells were cultured in 6-well plate at 37 °C and 5% CO2 for 7 days (M0). Macrophage polarization was obtained by removing the culture medium and culturing cells for an additional 48 h in RPMI 1640 supplemented with 5% FCS and 100 ng/ml LPS plus 20 ng/ml IFN-γ (for M1 polarization) or 20 ng/ml IL-4 (for M2 polarization).