A novel exvivo murine retina angiogenesis (EMRA) assay

Sara Rezzola, Mirella Belleri, Domenico Ribatti, Ciro Costagliola, Marco Presta, Francesco Semeraro

Research output: Contribution to journalArticlepeer-review


Pathological retinal angiogenesis results from the imbalance of pro-angiogenic and anti-angiogenic factors. In particular, vascular endothelial growth factor (VEGF) plays a pivotal role in retinal neovascularization and various therapeutic VEGF blockers have evolved over time. Nevertheless, new retinal angiogenesis models are crucial for investigating anti-angiogenic therapies and bringing them to patients. Here, we developed a novel exvivo murine retina angiogenesis (EMRA) assay in which endothelial sprouts originate from mature and quiescent retinal vessels. In this model, retina fragments from adult mice are embedded in a three-dimensional fibrin gel in the presence of human recombinant VEGF. Starting from the 3rd-4th day of incubation, endothelial cell sprouts invading the fibrin gel can be observed under an inverted microscope and measured at different time points thereafter. The effect of VEGF is dose-dependent, maximal stimulation being observed at day 7 for retina fragments stimulated with 25-75ng/ml of the growth factor. To assess whether the EMRA assay is suitable for testing the activity of anti-angiogenic compounds, retina fragments were incubated with VEGF in the presence of the neutralizing anti-VEGF antibodies bevacizumab and ranibizumab. The results demonstrate that both antibodies inhibit VEGF activity in a dose-dependent manner. In conclusion, the EMRA assay represents a new exvivo model of retinal neovascularization suitable for the rapid screening of novel anti-angiogenic therapeutics.

Original languageEnglish
Pages (from-to)51-56
Number of pages6
JournalExperimental Eye Research
Publication statusPublished - Jul 2013


  • Angiogenesis
  • Bevacizumab
  • Endothelium
  • Mouse
  • Ranibizumab
  • Retina
  • VEGF

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience


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