Research Article

Phenotypic and Functional Properties of Porcine Dedifferentiated Fat Cells during the Long-Term Culture In Vitro

Figure 4

Multilineage differentiation of porcine DFAT cells. (a) Adipogenic differentiation of DFAT cells at P3, P20, and P50 was induced by MDI (high-glucose DMEM containing 10% FBS, 1 μM dexamethasone, 500 μM isobutylmethylxanthine (IBMX), 10 μg/mL insulin, and 200 μM indomethacin) for 12 days. Lipid droplets were stained by Oil Red O, bar, 50 μm. Expression of PPARγ mRNA was assessed by real-time PCR. (b) Osteogenic differentiation of DFAT cells at P3, P20, and P50 was induced by induction medium (high-glucose DMEM containing 10% FBS, 0.1 μM dexamethasone, 10 mM β-glycerophosphate, and 50 mM ascorbic acid) for 21 days, bar, 200 μm. Expression of Runx2 mRNA was assessed by real-time PCR. (c) Myogenic differentiation of DFAT cells at P15 and P25 was induced by Galectin-1 for 21 days, expression of myogenic markers was confirmed by immunofluorescence analysis for Desmin and MyHC, and the nuclei were stained by DAPI. Images were merged by Desmin (red) and MyHC (green) with DAPI (blue), bar, 50 μm. Expression of MyoG mRNA was assessed by real-time PCR. All values are represented as mean ± SD from three independent experiments. , .
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