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Electrophoretically separated bone cell types from the foetal rat calvarium: A histochemical and biochemical study

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Summary

Isolated cells obtained from foetal rat bone (calvarium) by collagenase digestion can be separated into three subpopulations on the basis of surface charge by free flow electrophoresis. These subpopulations have been tentatively identified by numerical, biochemical and functional criteria and are believed to be composed of: (1) bone resorbing cell types, designated Peak I cells; (2) fibroblasts and loose connective tissue cells, designated Peak II cells; and (3) a mixture of osteoblasts and osteoprogenitor cell types, designated Peak III cells. the anatomical position of these subpopulations in the whole calvarium was determined by comparing the results of histochemical and morphological experiments with the results of biochemical experiments. It was found that Peak I cells are located predominantly on the ventral (endocranial) surface, Peak II cells in the connective tissue periosteal membranes and Peak III cells on the dorsal (ectocranial) surface and in the suture line areas. The response of these cell types to parathyroid hormone and calcitonin with regard to cAMP production and45Ca release from devitalized bone is examined and indicates that cells from Peak I and Peak III both respond to parathyroid hormone but only the cells from Peak I respond to calcitonin.

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References

  • Altman, A. J., Bandelin, J. G., Dominguez, J. H. &Mundy, G. R. (1978) Differentiation of isolated calvarial cells into a mature heterogenous bone cell population in culture.Met. Bone Dis. Rel. Res. 1, 75–9.

    Google Scholar 

  • Bessey, O., Lowry, O. &Brock, M. (1946) A method for the rapid determination of alkaline phosphatase.J. biol. Chem. 164, 321–9.

    Google Scholar 

  • Boyle, W. (1968) An extension of the51Cr-release assay for the estimation of mouse cytotoxins.Transplantation 6, 761–4.

    Google Scholar 

  • Burks, J. &Peck, W. A. (1978) Bone cells: a serum-free medium supports proliferation in primary culture.Science 199, 542–4.

    Google Scholar 

  • Chen, T. L. & Feldman, D. (1980) 1,25(OH)2D3 receptor concentration in primary bone cell cultures is altered by the rate of cell division.Am. Soc. Bone Min. Res. 34A (abstract).

  • Deshmukh, K. &Kline, W. G. (1976) Characterization of collagen and its precursors synthesized by rabbit articular cartilage cells in various culture systems.Eur. J. Biochem. 69, 117–23.

    Google Scholar 

  • Dziak, R. &Brand, J. S. (1974) Calcium transport in isolated bone cells: bone cell isolation procedures.J. Cell Physiol. 84, 75–84.

    Google Scholar 

  • Fishman, D. A. &Hay, E. D. (1962) Origin of osteoclasts from mononuclear leucocytes in regenerating newt limbs.Anat. Rec. 143, 329–37.

    Google Scholar 

  • Goldner, J. (1938) A modification of the Masson trichrome technique for routine laboratory purposes.Am. J. Pathol. 14, 237–42.

    Google Scholar 

  • Ham, A. W. (1974)Histology. 7th edn. Philadelphia: Lippincott.

    Google Scholar 

  • Hannig, K. (1972) Separation of cells and particles by continuous free flow electrophoresis. InTechniques of Biochemical and Biophysical Morphology (edited byGlick, D. andRosenbaum, R. M.), pp. 423–440. New York: Wiley.

    Google Scholar 

  • Hawker, C. D., Glass, J. D. &Rasmussen, H. (1966) Further studies on the isolation and characterization of parathyroid polypeptides.Biochemistry 5, 344–51.

    Google Scholar 

  • Jee, W. S. S. &Nolan, P. D. (1963) Origin of osteoclasts from the fusion of phagocytes.Nature, Lond. 200, 225–6.

    Google Scholar 

  • Kono, T. (1968) Purification and partial characterization of collagenolytic enzymes fromClostridium histolyticum.Biochemistry 7, 1106–14.

    Google Scholar 

  • Luben, R. A., Wong, G. L. &Cohn, D. V. (1976) Biochemical characterization with parathormone and calcitonin of isolated bone cells: Provisional identification of osteoclasts and osteoblasts.Endocrinology 99, 526–34.

    Google Scholar 

  • Mundy, G., Altman, A., Gondek, M. &Bandelin, J. (1977) Direct resorption of bone by human monocytes.Science 196, 1109–12.

    Google Scholar 

  • Peck, W. A., Birge, S. &Brandt, J. (1967) Collagen synthesis by isolated bone cells: Stimulation of ascorbic acidin vitro.Biochem. biophys. Acta 142, 512–25.

    Google Scholar 

  • Peck, W. A., Birge, S. A. &Fedak, S. A. (1964) Bone cells: Biochemical and biological studies after enzymatic isolation.Science 146, 1476–7.

    Google Scholar 

  • Puzas, J. E. &Brand, J. S. (1979) Parathyroid hormone stimulation of collagenase secretion by isolated bone cells.Endocrinology 104, 559–62.

    Google Scholar 

  • Puzas, J. E. &Goodman, D. B. P. (1978) A rapid assay for cellular deoxyribonucleic acid.Analyt. Biochem. 86, 50–5.

    Google Scholar 

  • Puzas, J. E., Vignery, A. &Rasmussen, H. (1979) Isolation of specific bone cell types by free flow electrophoresis.Calcif. Tiss. Intl. 27, 263–8.

    Google Scholar 

  • Randerath, K. (1966)Thin Layer Chromatography. 2nd edn. p. 110. New York: Academic Press.

    Google Scholar 

  • Rasmussen, H. &Bordier, P. (1974)The Physiological and Cellular Basis of Metabolic Bone Disease. Baltimore: Williams and Wilkins.

    Google Scholar 

  • Thompson, E. R., Baylink, D. J. &Wergedal, J. E. (1975) Increases in number and size of osteoclasts in response to calcium or phosphorus deficiency in the rat.Endocrinology 97, 283–94.

    Google Scholar 

  • Wong, G. L. &Cohn, D. V. (1975) Target cells in bone for parathormone and calcitonin are different: Enrichment of each type by sequential digestion of mouse calvaria and selective adhesion to a polymeric surface.Proc. natn. Acad. Sci., U.S.A. 72, 3167–71.

    Google Scholar 

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Puzas, J.E., Jensen, J.A. Electrophoretically separated bone cell types from the foetal rat calvarium: A histochemical and biochemical study. Histochem J 14, 561–571 (1982). https://doi.org/10.1007/BF01011889

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  • DOI: https://doi.org/10.1007/BF01011889

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