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In vitro and ex vivo retina angiogenesis assays

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Abstract

Pathological angiogenesis of the retina is a key component of irreversible causes of blindness, as observed in proliferative diabetic retinopathy, age-related macular degeneration, and retinopathy of prematurity. Seminal studies in the early 1980 s about the angiogenic activity exerted by mammalian retinal tissue extracts on the chick embryo chorioallantoic membrane and the later discovery of vascular endothelial growth factor (VEGF) accumulation in eyes of patients with diabetic retinopathy paved the way for the development of anti-angiogenic VEGF blockers for the treatment of retinal neovascularization. Since then, numerous preclinical and clinical studies about diabetic retinopathy and other retinal disorders have opened new lines of angiogenesis inquiry, indicating that limitations to anti-VEGF therapies may exist. Moreover, the production of growth factors other than VEGF may affect the response to anti-VEGF approaches. Thus, experimental models of retinal angiogenesis remain crucial for investigating novel anti-angiogenic therapies and bringing them to patients. To this aim, in vitro and ex vivo angiogenesis assays may be suitable for a rapid screening of potential anti-angiogenic molecules before in vivo validation of the putative lead compounds. This review focuses on the different in vitro and ex vivo angiogenesis assays that have been developed over the years based on the isolation of endothelial cells from the retina of various animal species and ex vivo cultures of neonatal and adult retina explants. Also, recent observations have shown that eye neovascularization in zebrafish (Danio rerio) embryos, an in vivo animal platform experimentally analogous to in vitro/ex vivo models, may represent a novel target for the identification of angiogenesis inhibitors. When compared to in vivo assays, in vitro and ex vivo models of retina neovascularization, including zebrafish embryo, may represent cost-effective and rapid tools for the screening of novel anti-angiogenic therapeutics.

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Acknowledgments

This work was supported in part by grants from Ministero dell’Istruzione, Università e Ricerca (MIUR, Centro IDET, FIRB project RBAP11H2R9 2011) and Associazione Italiana per la Ricerca sul Cancro (AIRC Grant No. 10396) to MP, European Union Seventh Framework Programme (FPT7/2007-2013) under Grant Agreement No 278570 to DR, and from Ministero dell’Istruzione, Università e Ricerca (MIUR, PRIN projects 32E9J9_002 and 32E9J9_004 2008) to CC and FS, respectively.

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Authors and Affiliations

  1. Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11, 25123, Brescia, Italy

    Sara Rezzola, Mirella Belleri, Giuseppina Gariano & Marco Presta

  2. Department of Ophthalmology, University of Brescia, Spedali Civili, Piazzale Spedali Civili 1, 25123, Brescia, Italy

    Sara Rezzola & Francesco Semeraro

  3. Unit of Human Anatomy and Histology, Department of Basic Biomedical Sciences, Neuroscience and Sensory Organs, University of Bari, Bari, Italy

    Domenico Ribatti

  4. National Cancer Institute, Giovanni Paolo II, Bari, Italy

    Domenico Ribatti

  5. Department of Medicine and Health Sciences, University of Molise, Campobasso, Italy

    Ciro Costagliola

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  1. Sara Rezzola
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Correspondence to Francesco Semeraro or Marco Presta.

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Rezzola, S., Belleri, M., Gariano, G. et al. In vitro and ex vivo retina angiogenesis assays. Angiogenesis 17, 429–442 (2014). https://doi.org/10.1007/s10456-013-9398-x

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