Porcine MSCs Adipogenic Differentiation

Porcine mesenchymal stem cells were isolated from adipose tissue and bone marrow from a 3-month-old Polish Large White pig, as described by Kociucka et al. (2016 (link)). The cells were cultured in advanced DMEM medium (Gibco) supplemented with 10% FBS (Sigma-Aldrich), 5 ng/ml FGF-2 (PromoKine), 2 mM L-Glutamine (PAA), 1 mM 2-mercaptoethanol (Sigma-Aldrich), 1 × antibiotic antimycotic solution (Sigma Aldrich), and MEM NEAA (Thermo Fisher Scientific) at 37 °C with 5% CO₂ supplementation. To induce adipogenic differentiation, the cells were grown to confluency and were cultured with adipogenic differentiation medium composed of advanced DMEM (Gibco) with 10% (v/v) FBS (Sigma), 1 × antibiotic antimycotic solution (Sigma Aldrich), MEM NEAA (Thermo Fisher), 5 ng FGF-2 (PromoKine), 1 × linoleic acid albumin (Sigma-Aldrich), 1 × ITS (Sigma-Aldrich), 1 μM dexamethasone (Sigma-Aldrich), 100 μM indomethacin (Sigma-Aldrich), and 50 mM IBMX (Sigma-Aldrich). The differentiation process was allowed to proceed for 7 days and lipid droplet formation was examined using a phase-contrast microscope (TS100 Eclipse, Nikon).