Fig. 6
BD treatment is sufficient to induce chondrogenic differentiation. The hMSCs were seeded at a density of 2 × 104 cells/cm2 in the absence or presence of chondral basal medium, CIM, and CIM + BD (100 μg/ml) for 14 days. Cells were fixed and stained with Alcian blue. a Represented images of chondrogenic differentiated cells. b The Alcian blue-stained GAGs were extracted, and their optic density was read at OD 620 nm. The results are presented as mean ± SEM from three independent experiments. *, P < 0.05, **, P < 0.01, ***, P < 0.005 vs. control. c The 3 × 105 cells were placed into a 15 ml conical tube, and centrifuged to yield micromass. Cells were treated in the absence or presence of chondral basal medium, CIM, and CIM + BD (100 μg/ml) for 14 days. The cell pellet in each tube was cryosectioned and stained with Alcian blue. d The Alcian blue stained GAGs were quantified using Image J software. *, represents P < 0.05 vs. related controls, as indicated. Scale bar: 100 μm. e hMSCs were seeded at a density of 2 × 104 cells/cm2 in the absence or presence of chondral basal medium, BD 100 (100 μg/ml), chon basal plus TGF-β with different doses (from 0.1–5 ng/ml), chon basal plus BD with various doses (from 1 to 500 μg/ml) for 7 days. Cells were fixed and stained with Alcian blue. f The Alcian blue-stained GAGs were extracted, and their optic density was read at OD 620 nm. The results are presented as mean ± SEM from three independent experiments. *, P < 0.05, **, P < 0.01, ***, P < 0.001 vs. control. Scale bar: 100 μm