Mouse AdMSCs Isolation and TGF-β1/Rosiglitazone Treatment

Mouse AdMSCs were isolated from adult male C57BL/6J mice’s inguinal fat pads as previously described (28 (link)). Briefly, the inguinal fat pads from 8- to 10-week-old male C57BL/6 mice were harvested, washed with PBS, and minced, followed by digestion in 10 mL of type I collagenase (1 mg/mL in 1% BSA/PBS) for 30 minutes at 37°C. The digested fat pads were filtered through a 40 μm cell strainer and centrifuged at 500g for 5 minutes at room temperature. The supernatant containing adipocytes and debris was discarded. Pelleted cells were resuspended and washed with PBS 2 times and used as AdMSCs. AdMSCs were cultured in DMEM/F12 (MilliporeSigma) supplemented with 10% fetal bovine serum (MilliporeSigma) and antibiotics (penicillin and streptomycin, Hyclone). AdMSCs were maintained at 37°C under a 5% CO₂ atmosphere. For the rosiglitazone treatment experiment, AdMSCs were treated with/without recombinant TGF-β1 (10 ng/mL) (Invitrogen) and rosiglitazone (10 μM) for 7 days after seeding (10 × 10⁴ cells/mL) in 6-well plates.