Human arterial smooth muscle cells in culture: inverse relationship between proliferation and expression of contractile proteins.
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, growth-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 stress 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 dedifferentiated 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 dedifferentiation 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.
Fager
G
G
Department of Medicine I, University of Göteborg, Sweden.
Hansson
G K
GK
Gown
A M
AM
Larson
D M
DM
Skalli
O
O
Bondjers
G
G
D000199:Actins / Q000032:analysis; D000941:Antigens / Q000032:analysis; D001158:Arteries; D001769:Blood; D002454:Cell Differentiation; D002455:Cell Division; D002467:Cell Nucleus / Q000276:immunology; D002478:Cells, Cultured; D003285:Contractile Proteins / Q000096:biosynthesis*; D003470:Culture Media; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D009218:Myosins / Q000032:analysis; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D014599:Uterus / Q000098:blood supply
D000199:Actins; D000941:Antigens; D003285:Contractile Proteins; D003470:Culture Media; D010982:Platelet-Derived Growth Factor; D009218:Myosins