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Ion-transporting ATPases and matrix mineralization in cultured osteoblastlike cells

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Summary

Cultures of osteoblastlike cells obtained from the endosteal surfaces of rabbit long bones formed and mineralized an extracellular matrix when they were supplied daily with medium containing fresh ascorbate. No matrix formed without this supplementation. The matrix mineralized whether or not beta-glycerophosphate, a substrate of alkaline phosphatase, was added to the medium. The ion-transporting ATPase activities of untreated, ascorbate-treated, and ascorbate plus beta-glycerophosphate-treated cells were measured. Ascorbate-treated and ascorbate plus beta-glycerophosphate-treated cells had similar enzyme activities. The activities of the Ca2+-ATPase; Ca2+,Mg2+-ATPase; and alkaline phosphatase in treated cells were elevated over the activities in untreated cells. Na+,K+-ATPase activity was lower in treated than in untreated cells. HCO3 −-ATPase activity was not changed by treatment. Alkaline phosphatase activity was 20 times higher in freshly isolated osteoblastlike cells than in cells grown to confluence in primary culture. In addition, subculturing further reduced the activity of this osteoblast-marker enzyme. The activities of the ion-transporting ATPases and alkaline phosphatase in second passage cells were similar to the activities of these enzymes in fresh, noncalcifying tissues. Nevertheless, second passage cells retain the ability to mineralize an extracellular matrix, and their ion-transporting ATPase and alkaline phosphatase activities are altered when the cells mineralize a matrix.

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Authors and Affiliations

  1. Division of Neuropharmacology and Epileptology, Department of Physiology, University of Utah, School of Medicine, 84108, Salt Lake City, Utah

    R. E. Anderson, J. W. Kemp & D. M. Woodbury

  2. Division of Radiobiology, Department of Pharmacology, University of Utah, School of Medicine, 84108, Salt Lake City, Utah

    W. S. S. Jee

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  1. R. E. Anderson
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  2. J. W. Kemp
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  3. W. S. S. Jee
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  4. D. M. Woodbury
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Additional information

This work was supported by Grant NAG-2-108 from the National Aeronautics and Space Administration, Washington, D.C., and Grant 5 PO1 NS15767 from the National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, MD.

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Anderson, R.E., Kemp, J.W., Jee, W.S.S. et al. Ion-transporting ATPases and matrix mineralization in cultured osteoblastlike cells. In Vitro 20, 837–846 (1984). https://doi.org/10.1007/BF02619629

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  • DOI: https://doi.org/10.1007/BF02619629

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