Figure 7.

Analysis of In Vivo Angiogenetic Potential of mRNA-Engineered EPCs in Chick Embryo Chorioallantoic Membrane Assay

(A) Schematic representation of the chorioallantoic membrane (CAM) assay. ANG-1, SDF-1α, and VEGF-A mRNA-transfected and untreated cells (medium or medium containing transfection reagent [TR]) were applied in silicone rings (8-mm inner diameter) onto the CAMs at the ninth day of incubation at 37°C and 60% humidity. At the fourth day of incubation, the CAMs were fixed and excised. (B) Analysis of the region of the inner ring circle with Wimasis WimCAM image software to quantify angiogenesis. Scale bar of photographs (upper) and analyzed pictures (lower) represents 2.7 mm. (C) Quantification of angiogenesis using Wimasis WimCAM web-based service. The vessel density, total branching points, total vessel network length, and total segments were quantified and compared to the medium control. Results are shown as mean + SD (n = 3). Statistical differences were determined using one-way ANOVA followed by Bonferroni multiple comparison test (**p < 0.01 and ***p < 0.001).