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Three-dimensional cellular development is essential for ex vivo formation of human bone

Nature Biotechnology volume 18, pages 954–958 (2000)Cite this article

Abstract

Tissue engineering of human bone is a complex process, as the functional development of bone cells requires that regulatory signals be temporally and spatially ordered. The role of three-dimensional cellular interactions is well understood in embryonic osteogenesis, but in vitro correlates are lacking. Here we report that in vitro serum-free transforming growth factor (TGF)-β1 stimulation of osteogenic cells immediately after passage results in the formation of three-dimensional cellular condensations (bone cell spheroids) within 24 to 48 hours. In turn, bone cell spheroid formation results in the up-regulation of several bone-related proteins (e.g., alkaline phosphatase, type I collagen, osteonectin) during days 3–7, and the concomitant formation of micro-crystalline bone. This system of ex vivo bone formation should provide important information on the physiological, biological and molecular basis of osteogenesis.

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Figure 1: Cell spheroid and microspicule formation and the immunocytochemical analysis.
Figure 2: TEM analysis of bone cell spheroids and microspicules.
Figure 3: Raman analysis of microspicules.
Figure 4: Bone protein expression and alkaline phosphatase activity of bone cell spheroids.
Figure 5: Density-dependent induction of bone protein expression.
Figure 6: Bone cell spheroid formation induces α-integrin chain expression.
Figure 7: Inhibition of bone-cell spheroid and microspicule formation.

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Acknowledgements

The authors thank Laurie McCauley and Renny Franceschi for careful reading of the manuscript. Supported, in part, by Grants AG 43460 and HL 59495 from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Pediatrics, University of Michigan, Ann Arbor, 48109, Michigan

    Sujata Kale & Michael William Long

  2. Department of Surgery, University of Michigan, Ann Arbor, 48109, Michigan

    Sybil Biermann

  3. Department of Chemistry, University of Michigan, Ann Arbor, 48109, Michigan

    Claire Edwards, Catherine Tarnowski & Michael Morris

  4. Comprehensive Cancer Center, University of Michigan , Ann Arbor, 48109, Michigan

    Sybil Biermann & Michael William Long

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  1. Sujata Kale
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Correspondence to Michael William Long.

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Kale, S., Biermann, S., Edwards, C. et al. Three-dimensional cellular development is essential for ex vivo formation of human bone. Nat Biotechnol 18, 954–958 (2000). https://doi.org/10.1038/79439

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