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Choroid Sprouting Assay to Study Ocular Microvascular Proliferation Ex Vivo
Overview
This video demonstrates the development of an ex vivo model of choroidal sprouting to study ocular microvascular angiogenesis. The assay involves culturing mouse eye explants containing the choroid tissue attached to the sclera and retinal pigment epithelium under specific conditions that promote choroidal microvascular sprouting.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Preparation
- Add 5 mL of Penicillin/Streptomycin (10000 U/mL) and 5 mL and 10 mL of commercially available supplements to 500 mL of complete classic medium with serum. Aliquot 50 mL of the medium initially.
NOTE: Do not return any medium back to the stock to avoid contamination. - Put an aliquot of the com.......
Representative Results
Figure 1: Schematic illustration showing choroid sprouting assay.
Eyes were first enucleated and cut circumferentially about 0.5 mm posterior to the limbus. The cornea, iris, lens, and vitreous were removed. Then a 1 mm cut was made from the edge of the eye cup towards the optic nerve. A band was then cut circumferentially about 1.0 mm posterior to the cut edge and the band and the peripheral regions of the complex were se.......
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Source: Tomita, Y., et al. An Ex Vivo Choroid Sprouting Assay of Ocular Microvascular Angiogenesis. J. Vis. Exp. (2020)
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