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Updated: Jun 17, 2025
Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension
Published on: January 23, 2015
JR5558 mice are a reliable model to investigate subretinal fibrosis
Yashar Seyed-Razavi1,2, So-Ra Lee3, Jiawen Fan3
1Save Sight Institute, Discipline of Ophthalmology, Sydney Medical School, The University of Sydney, Sydney, NSW, 2000, Australia. yashar.seyedrazavi@sydney.edu.au.
Abstract:
Subretinal fibrosis is a major untreatable cause of poor outcomes in neovascular age-related macular degeneration. Mouse models of subretinal fibrosis all possess a degree of invasiveness and tissue damage not typical of fibrosis progression. This project characterises JR5558 mice as a model to study subretinal fibrosis. Fundus and optical coherence tomography (OCT) imaging was used to non-invasively track lesions. Lesion number and area were quantified with ImageJ. Retinal sections, wholemounts and Western blots were used to characterise alterations. Subretinal lesions expand between 4 and 8 weeks and become established in size and location around 12 weeks. Subretinal lesions were confirmed to be fibrotic, including various cell populations involved in fibrosis development. Müller cell processes extended from superficial retina into subretinal lesions at 8 weeks. Western blotting revealed increases in fibronectin (4 wk and 8 wk, p < 0.001), CTGF (20 wks, p < 0.001), MMP2 (12 wks and 20 wks p < 0.05), αSMA (12 wks and 20 wks p < 0.05) and GFAP (8 wk and 12 wk, p ≤ 0.01), consistent with our immunofluorescence results. Intravitreal injection of Aflibercept reduced subretinal lesion growth. Our study provides evidence JR5558 mice have subretinal fibrotic lesions that grow between 4 and 8 weeks and confirms this line to be a good model to study subretinal fibrosis development and assess treatment options.
Insights
JR5558 mice develop fibrotic lesions mimicking subretinal fibrosis. This model allows non-invasive tracking and assessment of potential treatments for age-related macular degeneration complications.
Area of Science:
- Ophthalmology
- Regenerative Medicine
- Animal Models
Background:
- Subretinal fibrosis is a leading cause of vision loss in neovascular age-related macular degeneration.
- Existing mouse models often involve invasive procedures and tissue damage.
- A robust, non-invasive model is needed to study fibrosis development and test therapies.
Purpose of the Study:
- To characterize JR5558 mice as a model for studying subretinal fibrosis.
- To evaluate the temporal development and fibrotic nature of lesions in JR5558 mice.
- To assess the efficacy of Aflibercept in mitigating subretinal fibrosis progression.
Main Methods:
- Non-invasive fundus and optical coherence tomography (OCT) imaging to track lesion progression.
- Quantification of lesion area and number using ImageJ software.
- Histological analysis of retinal sections, wholemounts, and Western blotting for fibrotic markers.
Main Results:
- Subretinal lesions in JR5558 mice expanded significantly between 4 and 8 weeks, stabilizing by 12 weeks.
- Lesions exhibited fibrotic characteristics with involvement of Müller cells and increased fibronectin, CTGF, MMP2, αSMA, and GFAP.
- Intravitreal Aflibercept treatment effectively reduced subretinal lesion growth.
Conclusions:
- JR5558 mice provide a valuable, non-invasive model for subretinal fibrosis research.
- The model accurately recapitulates key fibrotic processes relevant to age-related macular degeneration.
- This model facilitates the evaluation of therapeutic interventions for subretinal fibrosis.
