Additional file 3: of Endothelial properties of third-trimester amniotic fluid stem cells cultured in hypoxia

Adipogenic and osteogenic differentiation of amniotic fluid stem cells from both trimesters. (A) Oil Red O staining (left images) (scale bar = 200 μm) and relative quantification (right graph) (expressed by ratio compared with control cells) (n = 5) performed at day 21 of differentiation. (B) Osteogenic differentiation was attained and calcium salt accumulation was detected by Von Kossa staining (top left images, brown-black deposits) (scale bar = 100 μm); calcium deposits from differentiating cells were also extracted and quantified at different time points (top right graph) (n = 3). Alkaline phosphatase activity was observed by colorimetric substrate modification (bottom left images) on differentiating cells at different time points. Alkaline phosphatase activity was also relatively quantified by measuring the substrate absorbance after reaction (bottom right graph) (expressed by ratio of differentiating over control cells, n = 3). **P < 0.01, ***P < 0.001. Ab antibody, ALP alkaline phosphatase. (JPEG 8213 kb)