Ability of hemopoietic cytokines to replace M-CSF in the induction of osteoclastic differentiation
| Preincubation . | Combinations of cytokines with RANKL . | TRAP-positive cells/coverslip . |
|---|---|---|
| GM-CSF (5 ng/mL) | GM-CSF (50 ng/mL) | 0 |
| IL-3 (1 ng/mL) | IL-3 (1 ng/mL) | 0 |
| IL-3 (1 ng/mL) | IL-3 (1 ng/mL) + GM-CSF (50 ng/mL) | 0 |
| SCF (5 ng/mL) | SCF (100 ng/mL) | 0 |
| SCF (5 ng/mL) | SCF (100 ng/mL) + FL (100 ng/mL) | 18 ± 7 |
| SCF (5 ng/mL) | SCF (100 ng/mL) + VEGF (100 ng/mL) | 0 |
| FL (5 ng/mL) | FL (100 ng/mL) | 39 ± 14 |
| FL (5 ng/mL) | FL (100 ng/mL) + SCF (100 ng/mL) | 43 ± 23 |
| FL (5 ng/mL) | FL (100 ng/mL) + IL-3 (1 ng/mL) | 31 ± 12 |
| FL (5 ng/mL) | FL (100 ng/mL) + VEGF (100 ng/mL) | 41 ± 14 |
| Preincubation . | Combinations of cytokines with RANKL . | TRAP-positive cells/coverslip . |
|---|---|---|
| GM-CSF (5 ng/mL) | GM-CSF (50 ng/mL) | 0 |
| IL-3 (1 ng/mL) | IL-3 (1 ng/mL) | 0 |
| IL-3 (1 ng/mL) | IL-3 (1 ng/mL) + GM-CSF (50 ng/mL) | 0 |
| SCF (5 ng/mL) | SCF (100 ng/mL) | 0 |
| SCF (5 ng/mL) | SCF (100 ng/mL) + FL (100 ng/mL) | 18 ± 7 |
| SCF (5 ng/mL) | SCF (100 ng/mL) + VEGF (100 ng/mL) | 0 |
| FL (5 ng/mL) | FL (100 ng/mL) | 39 ± 14 |
| FL (5 ng/mL) | FL (100 ng/mL) + SCF (100 ng/mL) | 43 ± 23 |
| FL (5 ng/mL) | FL (100 ng/mL) + IL-3 (1 ng/mL) | 31 ± 12 |
| FL (5 ng/mL) | FL (100 ng/mL) + VEGF (100 ng/mL) | 41 ± 14 |
Bone marrow cells were preincubated for 24 hours to remove stromal cells. Normally during this phase, the survival of osteoclastic precursors is supported by M-CSF, which was replaced in these experiments by a variety of cytokines. After 24 hours, nonadherent cells were removed, washed, and resuspended in the cytokines shown, together with RANKL (30 ng/mL). TRAP-positive cells were enumerated after 6 days of incubation and expressed as mean ± SEM (6 cultures per variable).