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  • Reliability of the mouse model of choroidal neovascularization induced by laser photocoagulation.

Reliability of the mouse model of choroidal neovascularization induced by laser photocoagulation.

Investigative ophthalmology & visual science (2014-09-11)
Stephen H Poor, Yubin Qiu, Elizabeth S Fassbender, Siyuan Shen, Amber Woolfenden, Andrea Delpero, Yong Kim, Natasha Buchanan, Thomas C Gebuhr, Shawn M Hanks, Erik L Meredith, Bruce D Jaffee, Thaddeus P Dryja
ABSTRACT

We attempted to reproduce published studies that evaluated whether the following factors influence choroidal neovascularization (CNV) induced by laser photocoagulation in murine retinas: small interfering RNA (siRNA), cobra venom factor, complement factors C3 and C5, and complement receptor C5aR. In addition, we explored whether laser-induced CNV in mice was influenced by the vendor of origin of the animals. Reagents or genotypes reported by others to influence CNV in this model were assessed using our standard procedures. Retrospective analyses of control or placebo mice in many experiments were done to evaluate whether the CNV area induced by laser photocoagulation varied according to vendor. Administration of the following agents did not have a substantial impact on the CNV induced by laser burns in mice: siRNA, low-molecular-weight inhibitor of the C5a receptor (PMX53), or cobra venom factor. Jackson Laboratory (JAX) mice lacking either C3 or C5 had increased neovascularization compared to non-littermate JAX wild-type controls. Taconic mice lacking C3 had reduced CNV compared to non-littermate Taconic wild-type control mice. A retrospective analysis of vehicle-treated wild-type C57BL/6 mice used as controls across 132 experiments conducted from 2007 to 2010 revealed that mice purchased from JAX or from Charles River produced less neovascularization than mice from Taconic. We present our recommended methods for conducting experiments with the mouse laser-induced CNV model to enhance reproducibility and minimize investigator bias.

MATERIALS
Product Number
Brand
Product Description

Dexamethasone, British Pharmacopoeia (BP) Assay Standard
Dexamethasone, European Pharmacopoeia (EP) Reference Standard
Dexamethasone for peak identification, European Pharmacopoeia (EP) Reference Standard
USP
Dexamethasone, United States Pharmacopeia (USP) Reference Standard
Supelco
Dexamethasone, Pharmaceutical Secondary Standard; Certified Reference Material
Dexamethasone for system suitability, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%
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Dexamethasone, powder, γ-irradiated, BioXtra, suitable for cell culture, ≥80% (HPLC)
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Dexamethasone, meets USP testing specifications
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Dexamethasone, ≥98% (HPLC), powder
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Dexamethasone, tested according to Ph. Eur.
Supelco
Dexamethasone, VETRANAL®, analytical standard