Calcified Tissue International

, Volume 34, Issue 1, pp 291–294 | Cite as

Formation of bone by isolated, cultured osteoblasts in millipore diffusion chambers

  • D. J. Simmons
  • G. N. Kent
  • R. L. Jilka
  • D. M. Scott
  • M. Fallon
  • David V. Cohn
Laboratory Investigations


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.

Key words

Bone formation Osteoblasts Osteoclasts Bone Induction Alkaline Phosphatase Cell Culture 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    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, 1957PubMedCrossRefGoogle Scholar
  2. 2.
    Peck, W. A., Birge, S. J., Fedak, S. A.: Bone cells: biochemical and biological studies after enzymatic isolation, Science146:1476–1477, 1964PubMedGoogle Scholar
  3. 3.
    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, 1973PubMedGoogle Scholar
  4. 4.
    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, 1974CrossRefPubMedGoogle Scholar
  5. 5.
    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, 1975PubMedGoogle Scholar
  6. 6.
    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, 1977PubMedGoogle Scholar
  7. 7.
    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, 1980PubMedGoogle Scholar
  8. 8.
    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, 1976PubMedCrossRefGoogle Scholar
  9. 9.
    Luben, R. A., Wong, G. L., Cohn, D. V.: Parathormonestimulated resorption of devitalised bone by cultured osteoclast-type bone cells, Nature265:629–630, 1977CrossRefPubMedGoogle Scholar
  10. 10.
    Scott, D. M., Kent, G. N., Cohn, D. V.: Collagen synthesis in cultured osteoblast-like cells, Arch. Biochem. Biophys.201:384–391, 1980CrossRefPubMedGoogle Scholar
  11. 11.
    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, 1979Google Scholar
  12. 12.
    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, 1954PubMedGoogle Scholar
  13. 13.
    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, 1971Google Scholar
  14. 14.
    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, 1968Google Scholar
  15. 15.
    Nade, S., Burwell, R. G.: Decalcified bone as a substrate for osteogenesis, J. Bone Joint Surg.59-B:189–196, 1977Google Scholar
  16. 16.
    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, 1970CrossRefPubMedGoogle Scholar
  17. 17.
    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, 1970Google Scholar
  18. 18.
    Burstone, M. S.: In: Enzyme Histochemistry and its Application in the Study of Neoplasms, pp. 275–276. Academic Press, New York, 1962Google Scholar
  19. 19.
    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, 1972Google Scholar
  20. 20.
    Friedenstein, A. J., Piatetsky-Shapiro, I. I., Petrakova, K. V.: Osteogenesis in transplants of bone marrow cells, J. Embryol. Exp. Morphol.16:381–390, 1966PubMedGoogle Scholar
  21. 21.
    Friedenstein, A. J.: Precursor cells of mechanocytes, Int. Rev. Cytol. 327–359, 1976Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • D. J. Simmons
    • 1
  • G. N. Kent
    • 2
  • R. L. Jilka
    • 2
    • 3
  • D. M. Scott
    • 2
  • M. Fallon
    • 4
  • David V. Cohn
    • 2
    • 3
  1. 1.Division of Orthopedic SurgeryWashington UniversitySt. Louis
  2. 2.Research LaboratoriesV.A. Medical CenterKansas City
  3. 3.University of Kansas Medical CenterKansas City
  4. 4.Department of PathologyJewish Hospital of St. LouisSt. LouisUSA

Personalised recommendations