Stempro chondrocyte differentiation basal medium

To qualify isolated cells from human BM as hMSCs, cells were induced to differentiation into adipogenic, osteogenic and chondrogenic lineages. Briefly, for adipogenic differentiation, 5x10³ hMSCs/cm² were exposed to Complete MesenCult Adipogenic Medium containing MesenCult MSC Basal Medium (Stemcell) and 10% Adipogenic Stimulatory Supplement (Stemcell) for 3 weeks. For osteogenic differentiation, 2x10⁵ cells/cm² incubated with Complete MesenCult Osteogenic Medium including MesenCult MSC Basal Medium, Osteogenic Stimulatory Supplement, β-Glycerophosphate, Dexamethasone, Ascorbic acid (all from Stemcell) for 5 weeks. For chondrogenic differentiation, 7.5x10⁶ cells/cm² were cultured for 3 weeks with Stempro Chondrocyte Differentiation Basal Medium (Gibco) containing 10% Stempro Chondrogenesis Supplement (Gibco). Standard histochemical staining methods were applied. Osteogenic, adipogenic and chondrogenic differentiation were confirmed by Toluidine Blue, Oil Red O, Alcian Blue staining, respectively.

For chondrogenic differentiation, the hMSC line was harvested at p3 and seeded into 24-well plates at a density of 7.5 × 10⁵ cells/cm². To stimulate chondrogenic differentiation, the culture medium was replaced with Stempro chondrocyte differentiation basal medium (Gibco) containing 10% Stempro chondrogenesis supplement (Gibco). After 21 days of cultivation, the chondrogenic pellet was stained with Alcian Blue.