Spheroid based angiogenesis assays for
the analysis of pro- and antiangiogenic factors
| This picture shows an example how the spheroid based angiogenesis assay can be used to identify pro- and antiangiogenic factors. Endothelial spheroids (1 d) were embedded into collagen gels containing the indicated factors (VEGF [50ng/ml], FGF-2 [30ng/ml], SU: Inhibitor of VEGF induced endothelial sprouting). After 48 h, in vitro angiogenesis was digitally quantitated by measuring the length of the sprouts that had grown out of each spheroid (ocular grid at 100x magnification) using the digital imaging software DP-Soft (Olympus, Germany) analyzing at least 10 spheroids per experimental group and experiment. Untreated control spheroids show a low baseline sprouting even in the presence of up to 20% FCS, reflecting their quiescent phenotype. VEGF and FGF-2 were known as proangiogenic factors which stimulate EC sprouting. SU was found to inhibit VEGF induced but not FGF-2 dependent sprouting. | |