Adipogenic differentiation kit

hMSCs were purchased from the American Type Culture Collection (ATCC) (PCS-500-011) and maintained in culture at 37°C and 5% CO₂ using the MesenCult proliferation kit (StemCell Technologies, Canada). At passage three, the cells were collected and grown in 6-well plates at 1.5 × 10⁴ cells/ml density using MesenCult medium containing the adipogenic differentiation kit (StemCell Technologies) to differentiate into adipocytes. The cells were treated with the standard substances — LA, ILA, and PLA —at concentrations ranging from 0.31 to 10 mM, and the culture medium was refreshed every 2 days until the hMSCs were matured. Subsequently, on day 21, the matured hMSCs were pretreated with PLA, ILA, and LA for 6 h before treatment with tumor necrosis factor-α (TNF-α) (10 ng/ml). The conditioned medium was collected after 24 h. To assess lipid accumulation level, cells were fixed in 10% formaldehyde in phosphate-buffered saline (PBS) and washed twice. Then, they were stained with Oil Red O (ORO) solution (Sigma-Aldrich) for 10 min and repeatedly rinsed with distilled water. Images of cells were captured at a 25× magnification using a digital camera attached to an inverse phase contrast microscope (Leica TP 1020, Germany).