Figure 5.
Analysis of Wound-Healing Capacity of mRNA-Engineered EPCs via Wound Scratch Migration Assay
1 × 105 murine EPCs were cultivated overnight, and then they were transfected with 1.6 μg ANG-1, 0.8 μg VEGF-A, or 0.5 μg SDF-1α mRNA or with an mRNA cocktail containing 1.6 μg ANG-1, 0.8 μg VEGF-A, and 0.5 μg SDF-1α mRNA. The next day, cells were detached, and 28,000 EPCs with or without mRNA transfection were seeded in each chamber of Culture-Insert 3 wells in μ-dishes. After 5 hr, when the cells completely attached and covered the surface, an open wound field was generated. Immediately after the generation of wound areas (0 hr) and after 12, 24, and 36 hr, phase-contrast images were taken and closed wound areas were calculated using Tscratch software. Scale bar represents 500 μm. Results are shown as mean + SD (n = 8). Statistical differences were determined using one-way ANOVA followed by Bonferroni multiple comparison test (*p < 0.05 and ****p < 0.0001).