Differentiation in serum-free versus FBS-medium
(A) Simplified schematic illustration of 2D adipocyte differentiation process. Bovine SVC cells were proliferated in either serum-containing growth medium (10% FBS; GM) or serum-free growth medium (SFGM). Adipogenesis was induced in a differentiation medium with 3%FBS or a defined animal component-free medium (DMAD). Media was changed 3 times. To assess the formation of adipocytes in response to the adipogenic cocktail in either serum or serum-free conditions, we performed fluorescent staining of the lipid droplets and nuclei, and quantified them with a high content analyzer ImageXpress Pico (HCA). Ros, rosiglitazone; and Dex, dexamethasone.
(B) Representative fluorescent images of cells on day 8 of differentiation with 3%FBS or DMAD, stained with BODIPY (green lipid droplets) and Hoechst (blue nuclei). Scale bar, 100 μm.
(C) Quantification of adipogenesis at day 8 and 12 with an HCA. Data are represented as mean ± standard deviation (SD); error bars represent the SD of 3 independent experiments using 3 donors. Statistical significance was determined with a two-way ANOVA; NS, not significant; ∗p < 0.05.