Summary: A physiologically relevant cellular model to assess angiogenesis.
Model Description
The inhibition of vascular endothelial cell growth factor A (VEGF) pathway in in vitro proliferation assay is an integral part of the potency assessment of VEGF inhibitor candidates for intraocular vascular diseases.
The primary bovine retinal endothelium cells (BRECs) are physiologically relevant cells to assess the angiogenesis.
Species | Primary bovine retinal endothelium cells (BRECs) |
Induction | rhVEGF165 |
Positive control | Aflibercept (Eylea®) at 0.01 to 30 nM |
Indicator and Read-outs | Indicator: resazurin (7-Hydroxy-3H-phenoxazin-3-one-10-oxide) dye is irreversibly reduced to highly fluorescent metabolite resorufin (7-Hydroxy-3H-phenoxazin-3-one); (Hakkarainen et al., 2016). Detection: After 2h exposure to resazurin, the amount of resorufin is measured at 560 nm (ex), 590 nm (em) by using Cytation 3 multi-mode reader (BioTek Instruments, Winooski, VT, USA); (Hakkarainen et al., 2016). |
Outcomes and Read-Outs
Imaging
Representative examples of assays on BRECs
References
- Yang J, Wang X, Fuh G, Yu L, Wakshull E, Khosraviani M, Day ES, Demeule B, Liu J, Shire SJ, Ferrara N, Yadav S. Comparison of binding characteristics and in vitro activities of three inhibitors of vascular endothelial growth factor A. Mol Pharm 2014,11(10):3421-3430. doi: 10.1021/mp500160v.
- Hakkarainen JJ, Reinisalo M, Ragauskas S, Seppänen A, Kaja S, Kalesnykas G. Acute cytotoxic effects of marketed ophthalmic formulations on human corneal epithelial cells. Int J Pharm. 2016;511(1):73-78. doi: 10.1016/j.ijpharm.2016.06.135