Fig. 4: SZP NSC CM induce less angiogenesis in vivo than Ctrl NSC CM.

a To perform the in vivo angiogenesis assay in the CAM of chicken embryos we added NSC 48 h CM on top of a bio cellulose scaffold. Scaffold was positioned over the CAM at embryonic day 8 (E8). After 4 days, vessels in the perimeter of the scaffold were counted. b Representative images of CAM vessels as indicated. Scaffold was filled with NEM as negative control, Ctrl NSC CM and SZP NSC CM. Scale bar = 1 cm. c Quantification of number of vessels in each condition at E12, represented as fold change in vessel number compared to E8. Data for each cell line (Ctrl #1, #2, #3 and SZP #1, #2, #3) is shown in a correlative order control and graphed as mean ± SD; * p < 0.05 and ***p ≤ 0.001 according to Kruskal–Wallis test. d, e Ctrl NSC CM was incubated with 100 µg/ml of bevacizumab (Bvz) to inhibit VEGF-A signaling. d Representative images of CAM vessels as indicated. Scaffold was filled with NEM, NEM plus 50 ng/ml VEGFA, NEM plus 50 ng/ml VEGFA with 100 µg/ml Bvz, Ctrl NSC CM, Ctrl NSC CM with 100 µg/ml Bvz or SZP NSC CM. e Quantification of number of vessels in each condition at E12, represented as fold change in vessel number compared to E8. Data is shown as mean ± SD; *p < 0.05 and ***p ≤ 0.001 according to Kruskal–Wallis test