To study the effect of cAMP on the generation of osteoclasts from their precursor cells, the bone marrow cells were isolated from 7 to 9 week-old male mice. The femur and tibia were dissected aseptically and the marrow cavity was flushed with 1
ml
of
α-minimum by slow injection. Collected marrow cells were cultured at 1.5-2.0×106 cells/well in 24-well plate for 8 days. In experimental group. PCE2(5×10-6, 10-5M), forskolin(10-5M) or IBMX(10-4, 10-5M) were added singly or in combination from
the
1st day of culture. After cultures. Staining for tartrate-resistant acid phosphatase(TRACP)-marker enzyme of osteoclast-was performed according to the modified method of Burstone. The TRACP-positive multinucleated cells(MNC). Which have 3 or more
nuclei, were counted.
@ES The observed results were as follows.
@EN 1. In control group, TRACP-positive mononuclear cells were present but no TRACP-positive multinucleated cells appeared.
2. PGE2(5×10-6M) or forskolin(10-5M) significantly stimulated the formation of TRACP-positive MNC. Moreover, forskolin potentiated the TRACP-positive MNC formation induced by PGE2 when added simultaneously.
3. IBMX(10-4, 10-5M), when added alone, significantly stimulated the formation of TRACP-positive MNC. However, IBMX(10-5M) added in combination of PGE2(10-5M) partially inhibited the TRACP-positive MNC formation induced by PGE2.
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