The murine macrophage cell line RAW 264.7 (TIB-71, ATCC) (originating from BALB/c mice cells transformed with Abelson leukaemia virus [43 (link)]) was used as a model for the induction of a pro-immune or anti-inflammatory response. For facility of passaging, cells were cultured normally as a semi-adherent culture in 10 cm2 suspension culture dishes at 37 °C, 5% CO2 and 80% humidity in 12 mL DMEM (#D0819-500ML, Sigma-Aldrich, Saint-Quentin-Fallavier, France) medium supplemented with 5% FBS 100 U/mL penicillin and 0.1 mg/mL streptomycin (#P0781-100ML, Sigma). Cells were passaged by gentle flushing of the culture medium to remove the cells, with cells then pelleted (200 g, 3 min, RT), resuspended in medium and counted (Section 2.2.1 ). Cells were re-seeded at 5 × 106 cells/mL or 10 × 106 cells/mL for 2 or 3 days of growth, respectively.
For experiments, RAW 264.7 cells were seeded the day before treatment at a density of 3 × 104 cells per well (100µL medium volume) in regular adherent 96-well culture plates (#655180, Greiner Bio-One, Les Ulis, France) or 3 × 105 cells per well (300µL medium volume) in 12-well plates (#665180, Greiner Bio-One, Les Ulis, France).
For experiments, RAW 264.7 cells were seeded the day before treatment at a density of 3 × 104 cells per well (100µL medium volume) in regular adherent 96-well culture plates (#655180, Greiner Bio-One, Les Ulis, France) or 3 × 105 cells per well (300µL medium volume) in 12-well plates (#665180, Greiner Bio-One, Les Ulis, France).
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