Cell and Tissue Research

, Volume 304, Issue 3, pp 371–376

Retroviral transduction of alveolar bone cells with a temperature-sensitive SV40 large T antigen

  • Vehid Salih
  • Jonathan C. Knowles
  • Michael J. O'Hare
  • Irwin Olsen
Regular Article

DOI: 10.1007/s004410100391

Cite this article as:
Salih, V., Knowles, J.C., O'Hare, M.J. et al. Cell Tissue Res (2001) 304: 371. doi:10.1007/s004410100391

Abstract.

We have transduced adult human alveolar bone (AB) cells with a gene construct encoding a temperature-sensitive mutation of the SV40 large T antigen (tsT). Such cells divided rapidly, for more than 50 passages thus far, at a permissive low temperature (34.5°C), comparable to the non-transduced parental cells at 37°C. However, the tsT-transduced AB cells failed to grow at a non-permissive high temperature (39°C) at which the T antigen is inactivated. Nevertheless, the cells formed mineralised nodules in vitro at both the low and high temperatures. Flow cytometry analysis showed that the transduced cells cultured at 34.5°C, like the parental cells at 37°C, were smaller and less granular than the transduced cells incubated at 39°C. Moreover, the transduced cells grown at 34.5°C were also found to express bone sialoprotein, osteopontin and type I collagen at levels similar to those of the parental cells at 37°C, although osteonectin and fibronectin were down-regulated. When the transduced cells were incubated at 39°C, the expression of all antigens was up-regulated, particularly osteonectin. Thus, we have obtained long-term cultures of tsT-transduced AB cells whose growth is temperature-dependent and which express certain features characteristic of bone-derived cells.

Osteoblasts Cell culture Flow cytometry Alveolar bone cells Human

Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  • Vehid Salih
    • 1
  • Jonathan C. Knowles
    • 1
  • Michael J. O'Hare
    • 2
  • Irwin Olsen
    • 3
  1. 1.Department of Biomaterials, Eastman Dental Institute, University College London, 256 Gray's Inn Road, London, WC1X 8LD, UK
  2. 2.Ludwig Institute of Cancer Research/Breast Cancer Laboratory, Department of Surgery, University College London Medical School, 67–73 Riding House Street, London, W1P 7LD, UK
  3. 3.Room RL 16, Department of Periodontology, Eastman Dental Institute, University College London, 256 Gray's Inn Road, London, WC1X 8LD, UK