In vitro differentiation of the murine macrophage cell line BDM-1 into osteoclast-like cells.
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.
Shin
J H
JH
Department of Microbiology, Saga Medical School, Japan.
Kukita
A
A
Ohki
K
K
Katsuki
T
T
Kohashi
O
O
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002117:Calcitriol / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D005260:Female; D007527:Isoenzymes / Q000378:metabolism; D008264:Macrophages / Q000166:cytology*; D008297:Male; D051379:Mice; D010010:Osteoclasts / Q000166:cytology*; D000071681:Tartrate-Resistant Acid Phosphatase; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007527:Isoenzymes; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase; D002117:Calcitriol; D013755:Tetradecanoylphorbol Acetate