Fig. 5

Analysis of angiogenesis in co-cultures of breast cancer cells and chemotherapy-exposed MSC using chorioallantoic model. (A) Outline of the experiment – MSC were exposed to the chemotherapy, co-cultured with tumor cells and topically applied on the CAM. (B) Evaluation of angiogenesis in co-culture of JIMT + MSC (unexposed or DOX-/PAC-exposed) on CAM by fractal analysis through quantification of fractal dimension (Df). We observed significantly higher vascular growth in DOX-MSC co-culture compared to unexposed MSC co-culture and JIMT only group. (C) Fractal analysis was also used to assess the vascularization in MDA co-culture with unexposed and chemotherapy-exposed MSC. Here we observed significantly higher vascular growth in both, DOX- and PAC-MSC co-cultures, compared to MDA only group. When compared to MDA + MSC, PAC-MSC co-culture had significantly more new blood vessels. (D) Co-cultures were growing on the CAMs in the area defined by a silicone ring. MDA cells co-cultured with DOX-MSC and PAC-MSC formed micro-metastases (indicated by arrows) outside that silicone ring. One-way ANOVA with Bonferroni multiple comparisons was used for statistical evaluation. Statistically significant results are highlighted with asterisks at p < 0.05 (*), p < 0.01 (**), p < 0.001 (***)