Kinetics of cell spreading in the presence of different concentrations of serum or fibronectin-depleted serum
A morphometric assay has been used to show that speed of cell spreading is sensitive to the level of serum in the surrounding growth medium. There was an inverse relationship between serum concentration and speed of spreading; as serum concentration was increased from 0.1% to 100% cells took progressively longer to become fully spread. The time taken to achieve complete spreading in serum concentrations of 0.1, 1.0, 10 and 100% serum was 45, 60, 105 and 240 min, respectively. At serum concentrations of 3% and above, fibronectin depletion had no effect on rates of spreading. Only at concentrations below 3% was there an effect of fibronectin depletion and although cells became fully spread in the depleted serum they took longer to do so in comparison to native serum. It is suggested that plasma fibronectin plays no role in serum-stimulated spreading at serum concentrations of 3% and above since at these concentrations rates of spreading are identical whether fibronectin is present or not. At concentrations of 3% and above spreading is mediated by the 70K spreading factor. At serum concentrations below 3% both fibronectin and 70K factor are effective. The reason for the lack of effect of fibronectin at higher serum concentrations is an inhibition by albumin and other plasma proteins. In contrast, 70K factor is not affected by exogenous protein concentration. The effect of exogenous protein concentration on cell adhesion is discussed in terms of cell migration between compartments that have different extracellular protein concentrations.