Endocrinology, Vol 136, 4285-4292, Copyright © 1995 by Endocrine Society

ARTICLES

In vitro differentiation of the murine macrophage cell line BDM-1 into osteoclast-like cells

JH Shin, A Kukita, K Ohki, T Katsuki and O Kohashi
Department of Microbiology, Saga Medical School, Japan.

Osteoclasts are derived from hematopoietic stem cells, but details about their precursor are still obscure. We present here a mouse macrophage cell line, BDM-1 cells, that showed a high potential to differentiate into osteoclast-like multinucleate cells (MNCs) when cocultured with primary osteoblasts for 14 days in the presence of 10(- 8) M 1 alpha,25-dihydroxyvitamin D3. These MNCs had tartrate-resistant acid phosphatase (TRAP) activity and strong ability to resorb dentine. In this culture system, 10(-10)-10(-8) M 12-O-tetradecanoylphorbol-13- acetate stimulated the formation of TRAP-positive MNCs, whereas salmon calcitonin inhibited it. Time-course effect studies showed that 12-O- tetradecanoylphorbol-13-acetate had an effect on the late phase of osteoclast differentiation but not on precursor proliferation. By immunocytochemical staining, all BDM-1 cells expressed Mac-1, Mac-2, and MOMA-2 antigens, and a large number of them expressed F4/80 antigen, but the rest of them were negative for this antigen. To select subclones able to differentiate into TRAP-positive MNCs, we sought to isolate several subclones from BDM-1 cells by mean of different specificity for F4/80 antigen expression. TRAP-positive MNCs were not generated from F4/80-positive subclones, but were obtained from subclones containing F4/80-negative cells. These results suggest that an F4/80-negative macrophage subpopulation is responsible for the differentiation of this cell line into osteoclasts.

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