Fig. 3

BD enhances CIM-induced chondrogenesis in high density cultures. The hMSCs were seeded at a density of 2 × 10 cells/cm2 for 14 days in the absence or presence of CIM with different dosages of BD (0, 0.1, 1, 10, 100, 500 μg/ml). a The morphological changes by phase contract microscopy. b The chondrogenic differentiation via Alcian blue staining. c The Alcian blue-stained glycosaminoglycans (GAGs) were extracted, and the absorbance was read at OD 620 nm. The results are presented as mean ± SEM from at least 5 independent experiments. *, P < 0.05, **, P < 0.01, ***, P < 0.001 vs. control. Scale bar: 100 μm. d hMSCs were seeded at 20000 cells/cm and treated with CIM in the absence or presence of BD (from 0.1-500 μg/ml) for 7 days. Cells were than harvested and the protein levels of SOX-9 and type II collagen were analyzed by Westerm blot. Cell lysate from untreated cells was used as control. e Quantification results of (d). The results are presented as mean ± SEM of two to three independent experiments. *, P < 0.05, **, P < 0.01 vs. control