Summary
Osteoblast-like and osteoclast-like cells freed from neonatal calvaria by sequential enzymatic digestion after 6–7 days in culture were placed in diffusion chambers and implanted in the peritoneal cavities of CD-1 mice. About half of the chambers also contained a dead calvarium to test for the need of an “inducer.” After 20 days, 11 of 18 chambers containing the osteoblast-like cells formed large foci of mineralized bone that corresponded to alkaline phosphatase activity throughout the chambers. Moreover, only type I (i.e., bone) collagen was formed. Occasional deposits of bone were found in only 3 of 22 chambers containing the osteoclast-like cells. The presence of dead bone did not affect any of the results. These data confirm the osteoblast-like nature of the isolated cell populations and demonstrate that these cells retain their differentiated function in culture.
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Jackson, S. F., Smith, R. H.: Studies on the biosynthesis of collagen. I. The growth of fowl osteoblasts and the formation of collagen in tissue culture, J. Biophys. Biochem. Cytol.3:897–912, 1957
Peck, W. A., Birge, S. J., Fedak, S. A.: Bone cells: biochemical and biological studies after enzymatic isolation, Science146:1476–1477, 1964
Peck, W. A., Carpenter, J., Messenger, K., De Bra, D.: Cyclic 3′5′-adenosine monophosphate in isolated bone cells: response to low concentrations of parathyroid hormone, Endocrinology92:692–697, 1973
Binderman, I., Duksin, D., Harell, A., Katzir, E., Sachs, L.: Formation of bone tissue in culture from isolated bone cells, J. Cell. Biol.61:427–439, 1974
Wong, G. L., Cohn, D. V.: Target cells in bone for parathormone and calcitonin are different: Enrichment for each cell type by sequential digestion of mouse calvaria and selective adhesion to polymeric surfaces, Proc. Natl. Acad. Sci. U.S.A.72:3167–3171, 1975
Peck, W. A., Burks, J. A., Wilkins, J., Rodan, S. B., Rodan, G. A.: Evidence for preferential effects of parathyroid hormone, calcitonin and adenosine on bone and periosteum, Endocrinology100:1357–1364, 1977
Williams, D. C., Boder, G. B., Toomey, R. E., Paul, D. C., Hillman, C. C., Jr., King, K. L., Van Frank, R. M., Johnston, C. C., Jr.: Mineralization and metabolic response in serially passaged adult rat bone cells, Calcif. Tissue Int.30:233–246, 1980
Luben, R. A., Wong, G. L., Cohn, D. V.: Biochemical characterization with parathormone and calcitonin of isolated bone cells: provisional identification of osteoclasts and osteoblasts, Endocrinology99:526–534, 1976
Luben, R. A., Wong, G. L., Cohn, D. V.: Parathormonestimulated resorption of devitalised bone by cultured osteoclast-type bone cells, Nature265:629–630, 1977
Scott, D. M., Kent, G. N., Cohn, D. V.: Collagen synthesis in cultured osteoblast-like cells, Arch. Biochem. Biophys.201:384–391, 1980
Cohn, D. V., Wong, G. L.: Isolated bone cells. In: D. J. Simons, A. S. Kanin (eds.): Skeletal Research—An Experimental Approach, pp. 3–20. Academic Press, New York, 1979
Algire, G. H., Weaver, J. M., Prehn, R. T.: Growth of cells in vivo in diffusion chambers; survival of homografts in immunized mice, J. Natl. Cancer Inst.15:493–507, 1954
Ostrowski, K., Wlodarski, K.: Induction of heterotropic bone formation. In G. H. Bourne (ed.): The Biochemistry and Physiology of Bone, Vol. III, Ed. 2, pp. 299–366. Academic Press, New York, 1971
Luna, L. G. (ed.): Manual of Histologic Staining of the Armed Forces Institute of Pathology, Ed. 3, pp. 94–95, 176–177. McGraw-Hill, New York, 1968
Nade, S., Burwell, R. G.: Decalcified bone as a substrate for osteogenesis, J. Bone Joint Surg.59-B:189–196, 1977
Huggins, C., Wiseman, S., Reddi, A. H.: Transformation of fibroblasts by allogeneic and xenogeneic transplants of demineralized teeth and bone, J. Exp. Med.132:1250–1258, 1970
Urist, M. R., Jurist, J. M., Dubuc, F. L., Strates, B. S.: Quantitation of new bone formation in intramuscular implants of bone matrix in rabbits, Clin. Orthop. Rel. Res.68:279–293, 1970
Burstone, M. S.: In: Enzyme Histochemistry and its Application in the Study of Neoplasms, pp. 275–276. Academic Press, New York, 1962
Pritchard, J. J.: The osteoblast. In G. H. Bourne (ed.): The Biochemistry and Physiology of Bone, Vol. I, pp. 21–43. Academic Press, New York, 1972
Friedenstein, A. J., Piatetsky-Shapiro, I. I., Petrakova, K. V.: Osteogenesis in transplants of bone marrow cells, J. Embryol. Exp. Morphol.16:381–390, 1966
Friedenstein, A. J.: Precursor cells of mechanocytes, Int. Rev. Cytol. 327–359, 1976
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Simmons, D.J., Kent, G.N., Jilka, R.L. et al. Formation of bone by isolated, cultured osteoblasts in millipore diffusion chambers. Calcif Tissue Int 34, 291–294 (1982). https://doi.org/10.1007/BF02411253
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DOI: https://doi.org/10.1007/BF02411253