Effects of immobilized Delta-1–FL on the differentiation into osteoclasts and dendritic cells of BM cells precultured with M-CSF.

BM cells cultured with M-CSF for 4 days were harvested, and adherent and nonadherent cells were further cultured with M-CSF (50 ng/mL) and RANKL (25 ng/mL) in wells coated with (A) Delta-1–FL or BAP-FL (2 μg), or (C) various doses of Delta-1–FL (2, 1, 0.5, 0.05, and 0 μg). (B) After 6 days, TRAP staining was performed. Adherent and nonadherent cells were cultured with GM-CSF (100 U/mL) and IL-4 (25 ng/mL) in the wells coated with Delta-1–FL or BAP-FL (2 μg) for 6 days, and the harvested cells were stained with anti-I-A^band anti-CD11c antibodies, and analyzed by flow cytometry. The numbers of I-A^b-high and CD11c⁺ cells were calculated as follows: recovered cell number/well × % positive cells/100. Bars indicate means ± SD of triplicate cultures. *Indicates significantly different from the corresponding cultures in dishes coated with BAP-FL (P < .01).