Human arterial smooth muscle cells in culture: Inverse relationship between proliferation and expression of contractile proteins |
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Authors: | Gunnar Fager Göran K Hansson Allen M Gown David M Larson Omar Skalli Göran Bondjers |
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Affiliation: | 1. Arterial Biology Group, Wallenberg Laboratory for Cardiovascular Research, Department of Medicine I, University of G?teborg, S-413 45, G?teborg, Sweden 2. Department of Clinical Chemistry, University of G?teborg, S-413 45, G?teborg, Sweden 3. Department of Pathology, University of Washington, Seattle, Washington 4. Cardiovascular Pathology, Mallory Institute of Pathology, Boston University School of Medicine, Boston, Massachusetts 5. Department of Pathology, University of Geneva, Geneva, Switzerland
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Abstract: | Summary Human arterial smooth muscle cells (hASMC) from explants of the inner media of uterine arteries were studied in secondary
culture. We had previously found that these cells depend on exogenous platelet-derived growth factor (PDGF) for proliferation
in vitro. Deprivation of the serum mitogen(s) by culture in plasma-derived serum or bovine serum albumin (BSA) caused a true
growth arrest that was reversible upon reexposure to the mitogen(s). When added to serum-containing medium, heparin caused
a reversible growth arrest which could be competed for by increasing concentrations of serum. In the current study we used
a set of smooth muscle-specific actin and myosin, antibodies to study the expression of contractile proteins in stress fibers
under indirect immunofluorescence on hASMC in culture. Even in sparse culture, grwoth-arrested hASMC expressed stress fibers
containing these actin and myosin epitopes. This was true irrespective of whether growth arrest was achieved by culture in
media containing only BSA or a combination of heparin and whole blood serum. hASMC proliferating in whole blood serum in sparse
culture did not express such strees fibers, as judged by immunofluorescent staining. This was true also for cells that were
restimulated to proliferate in serum after a growth arrest. Utilizing a monoclonal antibody against a nuclear antigen expressed
in proliferating human cells, we were able to demonstrate an inverse relationship between the expression of this antigen and
the SMC-specific contractile proteins, respectively. Under these culture conditions, the reversible transition between defifferentiated
and differentiated hASMC was almost complete and terminated about 1 wk after the change in culture condition. We conclude
that hASMC in vitro respond, to exogenous PDGF by proliferation and dedifferetiation as a single population of cells. We also
conclude that this modulation is reversible, because the cells become uniformly quiescent and differentiated when the mitogenic
stimulus is blocked or removed.
This study was supported by grants from the Swedish Medical Research Council (Project no. 4531 and 6816), the Swedish Association
against Heart and Chest Diseases, the King Gustaf V and Queen Victoria Foundation, the National Institutes of Health, Bethesda,
MD (grant HL 29873) and the Swedish National Board for Laboratory Animals. |
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Keywords: | smooth muscle cells human vascular differentiation cytoskeleton actin myosin |
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