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Proliferative responses to estradiol, IL-1α and TGFβ by cells expressing alkaline phosphatase in human osteoblast-like cell cultures

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Summary

The use of primary (nontransformed) bone cell cultures is hampered by their cellular heterogeneity. Primary cultures of osteoblast-like cells have been shown to proliferate in response to several osteotropic agents, but because mixed cell populations are present it is uncertain whether a true osteoblastic response was observed. By combining (1) localization of [3H]-thymidine incorporation into the nuclei of actively dividing cells by autoradiography with (2) subsequent induction of osteoblast differentiation by 1,25(OH)2D3 to optimize the number of cells expressing high alkaline phosphatase activity and (3) its localization by histochemical staining, it is possible to measure the proliferation of cells that are capable of expressing a more mature osteoblastic phenotype in heterogeneous human trabecular bone cell cultures.

Over a 72-hour incubation period, rhIL-1α (0.2–2 ng/ml) exerted a dose-dependent stimulation of proliferation of cells expressing alkaline phosphatase. Purified human TGFβ1 produced a biphasic increase in the proliferation of these cells (0.01–1 ng/ml) but 17β and 17α-estradiol (10-12–10-8 M) failed to consistently regulate cell growth. Furthermore, 17β-estradiol did not reproducibly modulate proliferation induced by IL-1α or TGFβ when added together in cultures. This procedure represents a more accurate method for the assessment of osteoblast proliferation in primary bone cell cultures and demonstrates that estrogen is not mitogenic for human osteoblasts and does not potentiate the actions of putative local stimulators of osteoblast replication.

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Author notes

  1. Maxine Gowen

    Present address: Department of Cellular Biochemistry, Smith-Kline Beecham Pharmaceuticals, 19406, King of Prussia, PA, USA

  2. Brian R. MacDonald

    Present address: Division of Rheumatology, Hospital of the University of Pennsylvania, 19104, Philadelphia, PA, USA

Authors and Affiliations

  1. Bath Institute for Rheumatic Diseases, Bath, Avon, UK

    David J. Rickard, Maxine Gowen & Brian R. MacDonald

  2. University of Bath, Bath, Avon, UK

    David J. Rickard, Maxine Gowen & Brian R. MacDonald

Authors
  1. David J. Rickard
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  2. Maxine Gowen
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  3. Brian R. MacDonald
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Rickard, D.J., Gowen, M. & MacDonald, B.R. Proliferative responses to estradiol, IL-1α and TGFβ by cells expressing alkaline phosphatase in human osteoblast-like cell cultures. Calcif Tissue Int 52, 227–233 (1993). https://doi.org/10.1007/BF00298724

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  • DOI: https://doi.org/10.1007/BF00298724

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