Fig. 5

Representative images of osteogenic, adipogenic, and chondrogenic differentiation assays as well as colony forming unit-fibroblast (CFU-F) assays performed on stromal vascular fraction (SVF) cells before freezing or after 2 months of storage in the presence of cryopreservation solutions (Sol.) C and D. Differentiation was induced (Stim.) by the addition of commercially available osteogenic, adipogenic, and chondrogenic differentiation media to cells at passage P1. Unstimulated cells (Unst.; control) were cultured with 10% FBS medium. Within the chondrogenesis assay, spheroids failed to be obtained from unstimulated cells. Differentiated cells as well as unstimulated samples were stained with Alizarin Red, Oil Red-O, and Safranin-O to detect osteocytes, adipocytes, and chondrocytes, respectively. The differentiation degree was quantified by image analysis of cell staining (adipogenesis and osteogenesis) or by morphometric analysis of spheroids (chondrogenesis); results are reported in histograms. Sample storage in the presence of solutions C and D did not significantly affect cell differentiation potential. Scale bar = 100 μm. C.A., covered area; Vol., volume