Stempro osteogenic

mSG-SCs were induced with 3 different culture media (CCM vs StemMacs vs Stem-Pro) towards osteogenic, chondrogenic, and adipogenic phenotype to evaluate their differentiation capacity at p.2–3, p.6–7, and p.10–11. For osteogenic and chondrogenic differentiation, 300,000 cells were seeded in 6 well plates until reaching confluency, and the medium was changed (StemPro osteogenic and StemPro chondrogenic differentiation kits; Life Technologies). After 14 days of continuous culture, the RNA of the cells was isolated to perform qPCR as described in previous section, and the expression of different osteogenic markers (ALP, BMP-2, bone gamma-carboxyglutamate protein/osteocalcin (BGLAP), and chondrogenic (ACAN, SOX-9)) was analyzed. Furthermore, mSG-SCs were evaluated for bone formation potential. Therefore, the wells were exposed to osteogenic medium for 21 days and further were evaluated by histochemical staining with 1% red of Alizarin (Alizarin Red S) followed by measurement of mineralized tissue by its dissolution coupled staining with cetyl pyridinium chloride (CPC), as already described previously (Andreadis et al. 2014 (link)). The optical density (OD) was measured at 550 nm with an ELISA spectrophotometer (Epock; Biotek Instruments, USA).