Figure 5.

Morphology and motility of HUVECs seeded on fibronectin, tenascin-W, or tenascin-C in combination with type I collagen. A) Culture plates were coated with type I collagen, followed by fibronectin (FN), tenascin-W (TNW) or tenascin-C (TNC). HUVECs were seeded on these mixed substrata and incubated for 3 h at 37°C. Cells were then fixed and stained with crystal violet, and morphology of the cells was observed. Morphology of HUVEC populations observed on the different substrata is shown. Note the emergence of elongated cells in the presence of TNW and TNC. Scale bar = 50 μm. B) Percentage of elongated cells on total number of adherent cells in each condition described in A. Statistical analyses were performed with ANOVA. C) Culture plates were coated with type I collagen. HUVECs were then seeded in serum-free medium containing increasing concentrations of soluble TNW or BSA as a control. After 3 h incubation at 37°C, cells were fixed, stained with crystal violet and analyzed. For each concentration of TNW, the number of HUVEC elongated cells was assessed, and the percentage of the total number of cells was calculated. Background, evaluated as percentage of HUVEC elongated cells observed in control conditions (i.e., BSA at the same concentration), was then deduced. D) Culture plates were coated with type I collagen, followed by FN, TNW, or TNC. After 2 h of incubation at 37°C, the movement of HUVECs was recorded by time-lapse microscopy. Cell motility was quantified, tracking ≥35 cells/condition throughout the time stack. Results are represented with a box plot; thicker line represents median value. Ten percent of the values are composed within the notch surrounding the median line. Bottom and top extensions of the box identify the first and the fourth quartile, respectively. Extreme lines represent the minimal and the maximal values of each dataset. Outliers are individually plotted as small circles. Note the significant increase (P<0.0001, ANOVA test) of cell motility in the presence of TNW and TNC.