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Characterization of vascular smooth muscle cell phenotype in long-term culture

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Summary

Studies of bovine carotid artery smooth muscle cells, during long-term in vitro subcultivation (up to 100 population doublings), have revealed phenotypic heterogeneity among cells, as characterized by differences in proliferative behavoir, cell morphology, and contractile-cytoskeletal protein profiles. In vivo, smooth muscle cells were spindle-shaped and expressed desmin and alpha-smooth muscle actin (50% of total actin) as their predominant cytoskeletal and contractile proteins. Within 24 h of culture, vimentin rather than desmin was the predominant intermediate filament protein, with little change in alpha-actin content. Upon initial subcultivation, all cells were flattened and fibroblastic in appearance with a concommitant fivefold reduction in alpha-actin content, whereas the beta and gamma nonmuscle actins predominated. In three out of four cell lines studied, fluctuations in proliferative activity were observed during the life span of the culture. These spontaneous fluctuations in proliferation were accompanied by coordinated changes in morphology and contractile-cytoskeletal protein profiles. During periods of enhanced proliferation a significant proportion of cells reverted to their original spindle-shaped morphology with a simultaneous increase in alpha-actin content (20 to 30% of total actin). These results suggest that in long-term culture smooth muscle cells undergo spontaneous modulations in cell phenotype and may serve as a useful model for studying the regulation of intracellular protein expression.

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Authors and Affiliations

  1. College of Medicine, Department of Physiology and Biophysics, University of Vermont, 05405, Burlington, Vermont

    Jan Woodcock-Mitchell, John Mitchell, Linda Baldor, Robert Low & David Warshaw

  2. College of Medicine, Department of Medicine, University of Vermont, 05405, Burlington, Vermont

    Marlene Absher

Authors
  1. Marlene Absher
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  2. Jan Woodcock-Mitchell
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  3. John Mitchell
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  4. Linda Baldor
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  5. Robert Low
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  6. David Warshaw
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Additional information

This work was supported by grants from from National Institutes of Health, Bethesda, MD, to DMW (HL35684), JW (HL36412), and JM and RL (SCOR HL 14212).

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Absher, M., Woodcock-Mitchell, J., Mitchell, J. et al. Characterization of vascular smooth muscle cell phenotype in long-term culture. In Vitro Cell Dev Biol 25, 183–192 (1989). https://doi.org/10.1007/BF02626176

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  • DOI: https://doi.org/10.1007/BF02626176

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