Fig. 10

Marked suppression of osteoblast-mediated calcification by cross-linking of A7 antigen without affecting cell proliferation Primary calvarial osteoblasts were cultured in 96 multi-well plates (3.2 × 10³ cells/well) as described in Materials and Methods. a A7 MAb markedly inhibited calcification mediated by primary osteoblasts. Primary osteoblasts were cultured in calcification medium as described in Materials and Methods in the absence (upper panels) or in the presence of indicated concentrations of A7 MAb (middle panels) or isotype control IgG (lower panels). At day 21, cells were fixed and stained using alizarin red for the assessment of calcified deposits formation. b A7 MAb has no effect on cell proliferation and viability of primary osteoblasts. Calvarial osteoblasts were cultured in α-MEM, 10% FBS in the presence of indicated concentrations of A7 MAb (left panel) or isotype control IgG (right panel). MTT proliferation assay was performed at day 4 culture. Data represent mean ± SEM (n = 4). Experiments were performed two times and obtained almost identical results