ADSC-EVs Regulate Macrophage Polarization

To investigate the role of ADSC-EVs in regulating macrophage polarization in an inflammatory environment, 5 × 10⁵ raw 264.7 cells were seeded in 6-well plates. After cell attachment, the culture medium was replaced with culture medium (control group), culture medium containing 1 mg/mL LPS (Sigma-Aldrich, St. Louis, MO, USA) and 20 ng/mL IFN-γ (PeproTech, Rocky Hill, USA) (LPS + IFN-γ group), or culture medium containing 1 mg/mL LPS, 20 ng/mL IFN-γ, and different concentrations of EVs (LPS + IFN-γ + EV group), as previously described [23 (link)]. After incubating the different media for 24 h, the polarization of raw 264.7 cells to M1/M2 macrophages was identified by flow cytometry, immunofluorescence staining, and quantitative real-time polymerase chain reaction (qPCR).