Human vascular smooth muscle cells in culture: growth characteristics and protein pattern by use of serum-free media supplements.
This study demonstrates that cultivation of vascular smooth muscle cells from human artery wall is possible under completely serum-free conditions. The effects of attachment factors on cell spreading and cell proliferation are described in detail as well as routine cultivation methods under serum-free conditions (clone cultures, cell migration, subcultivation by use of an exogenous trypsin inhibitor, cryopreservation and readaptation of cells). After a careful adaptation period, only two (BMS and Ultroser G) of the four commercially available serum-free media supplements tested were used successfully for a routine cultivation of the smooth muscle cells over several passages. With both supplements cell proliferation rates were comparable with those obtained in medium containing 10% fetal calf serum. The addition of platelet-derived growth factor or transferrin to serum-free cultures had no growth-stimulating effect. The addition of endothelial cell growth factor isolated from bovine brain caused a significant increase in proliferative activity of cells cultivated with BMS, but not with Ultroser G. Moreover, we report that under the serum-free culture conditions described here, the gamma-actin content of the cells is largely reduced (51% +/- 13% (means +/- SD) for cells cultivated in Ultroser G, and 12% +/- 4% (means +/- SD) for cells cultivated in BMS) when compared with cells cultivated under serum-containing conditions (gamma-actin content = 100%). The alpha-actin content was observed to be unaltered. Even after a careful readaptation of serum-free cultured cells to serum conditions, the gamma-actin content remained reduced.
Dartsch
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PC
Physiologisches Institut I der Universität, Tübingen/Bundesrepublik Deutschland.
Weiss
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HD
Betz
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D000199:Actins / Q000032:analysis; D002448:Cell Adhesion; D002455:Cell Division; D002465:Cell Movement; D002470:Cell Survival; D002478:Cells, Cultured / Q000502:physiology*; D003470:Culture Media; D015180:Electrophoresis, Gel, Two-Dimensional; D016228:Endothelial Growth Factors; D006133:Growth Substances / Q000502:physiology; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D010982:Platelet-Derived Growth Factor / Q000502:physiology; D011506:Proteins / Q000032:analysis; D014168:Transferrin / Q000502:physiology
D000199:Actins; D003470:Culture Media; D016228:Endothelial Growth Factors; D006133:Growth Substances; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D014168:Transferrin