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Fig. 5. Inhibition of adipogenesis of Runx2/ chondrocytes by soluble differentiation factors. Runx2/ primary chondrocytes were plated at a density of 1x105 cells/well (5x104 cells/cm2) in 24 multi-well plates. Two days after inoculation, the cells were further cultured for 10 days without (control) or with a factor including TGF-ß (1 ng/ml), retinoic acid (RA) (108 M), IL-1ß (10 ng/ml), PDGF (200 ng/ml), bFGF (10 ng/ml), IL-11 (100 ng/ml), PTH (107 M), BMP-2 (50 ng/ml), IGF-1 (100 ng/ml) or T3 (107 M), and stained with oil red O or Alcian blue. After culture, the cells were replated for the examination of type II collagen expression by immunocytochemistry. Magnification: phase contrast, oil red and Alcian blue, x50; type II collagen, x100.