In vitro model: Evercyte has generated telomerized human umbilical vein endothelial cells (HUVEC/TERT2) that can be grown without limitation while maintaining a primary-like phenotype. Assay set-up: HUVEC/TERT2
cells are seeded onto matrigel matrix and the formation of tubule-like structures is monitored. Additionally, HUVEC/TERT2 cells can be grown as 3D spheroids. Treatment of these spheroids with vascular endothelial growth factor (VEGF) leads to formation of sprouts indicating induction of neo-angiogenesis. The length and number of sprouts that are formed allows the identification of compounds with pro- or anti-angiogenic activities. Fast screening of compounds for neo-angiogenic potential is done by seeding HUVEC/TERT2 cells into multi-well plates followed by treatment with test compounds and measurement of cellular proliferation.Timelines:
depending on the bioassay, the number of samples, replicates and concentrations tested, it takes between 2 and 6 weeks to deliver results.
Testing neo-angiogenic potential of compounds
sprout formation from endothelial spheroids
When human umbilical vein endothelial cells (HUVEC/TERT2) are grown in hanging drops, they form 3D spheroids that can be embedded into a collagen matrix. Treatment of these spheroids with vascular endothelial growth factor (VEGF) induces the formation of sprouts. Concommitant cultivation in the presence of VEGF and Cyclosporin A (CSA) significantly reduces sprout formation in a concentration-dependent manner.
Testing endothelial cell proliferation
2D proliferation assay
Telomerized human endothelial cells (HUVEC/TERT2) are seeded into multi-well plates followed by treatment with vascular endothelial growth factor (VEGF) with / without anti-angiogenic agents ranibizumab (Lucentis®) and bevacizumab (Avastin®). Measurement of cellular proliferation demonstrates that Lucentis and Avastin inhibit VEGF induced proliferation in a concentration dependent manner.