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Department of Bone Biology and Osteoporosis, Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania 19486 [S. B. R., Y. I., M. A. T., G. W., D. T., G. A. R.]; The Hebrew University, Hadassah Medical School, Jerusalem, Israel [Z. B-S.]; and Department of Biochemistry, University of Vermont, Burlington, Vermont 05405 [S. S., K. M.]
This study examines the osteoblastic properties of the established human osteosarcoma cell line Saos-2. Saos-2 cells inoculated into diffusion chambers, which were implanted i.p. into nude mice, produced mineralized matrix in 4 of 6 chambers at 8 weeks. In 5 of 6 chambers there was a strong positive alkaline phosphatase reaction. In culture the alkaline phosphatase levels increased with time and cell density, reaching very high levels at confluence: 4–7 µmol/mg protein/min. The cells show a sensitive adenylate cyclase response to parathyroid hormone, 50% effective dose = 2.8 nM, which increases with cell density and is further raised by dexamethasone treatment. They also exhibit typical binding of 1-25-dihydroxyvitamin D3 to 3.2S receptor protein with an apparent Kd of 0.21 nM; the numbers of sites per cell were 3,300 at 50,000 cells/cm2 and 1,800 at 280,000 cells/cm2. The presence of osteonectin was visualized with a monoclonal antibody which revealed a reticular pattern on the cell surface. Osteonectin was also detected in the medium by Western blots, migrating at around Mr 40,000 in nonreduced gels and Mr 44,000 in reduced gels. The Saos-2 cells thus possess several osteoblastic features and could be useful as a permanent line of human osteoblast-like cells and as a source of bone-related molecules.
1 To whom requests for reprints should be addressed, at the Department of Bone Biology and Osteoporosis, Merck Sharp & Dohme Research Laboratories, West Point, PA 19486.
Received 3/ 6/87. Revised 6/ 5/87. Accepted 6/10/87.
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