Isolation and Characterization of Mouse Adipose-Derived SVFs

As above mentioned, SVFs were isolated from mouse adipose tissues. Flow cytometry was conducted as described^{51 (link)}. The SVFs were then stained in cold PBS containing 0.5% BSA and 2 mM EDTA (pH 7.4) with PE anti-mouse F4/80 (123110), FITC anti-mouse CD11b (101206) and APC anti-mouse CD11c (117310) for 30 min at 4 °C. After fixed with Fixation Buffer (420801) at 4 °C for 30 min, the cells stained in Permeabilization Wash Buffer (421002) with PE/Cy7 anti-mouse CD206 (141720) at 4 °C for 30 min. All the antibodies and reagents were purchased from Biolegend, San Diego, CA, USA. The antibody dilution was 1/400. The cells were subjected to flow cytometry analysis using a MACSQuantTM (Miltenyi Biotec, Auburn, CA, USA). Data analysis was performed using the FlowJo software (Tree Star Inc, Ashland, USA) as instructed.