Figure 3

The number of TRACP+ multinuclear cells. Mouse (C57BL/6, 8–12 weeks) bone marrow cells were isolated from mouse long bones and seeded in 24-well plates on bovine bone slices. Cells on the bone slices were cultured in four groups: control, VEGF (100 ng/ml) and ENDO (2 μg/ml) and VEGF + ENDO respectively. The control group was cultured in α-MEM with 10% FCS containing 30 ng/ml RANKL and 10 ng/ml M-CSF for 7 days, after which the cultures were stopped by fixing the cells with 3% paraformaldehyde (PFA)/2% sucrose in PBS.