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Updated: Jul 25, 2025
Characterization of a Novel Human Organotypic Retinal Culture Technique
Published on: June 9, 2021
Mammalian Animal and Human Retinal Organ Culture as Pre-Clinical Model to Evaluate Oxidative Stress and Antioxidant
Martina Kropp1,2, Mohit Mohit1,2, Cristina Ioana Leroy-Ciocanea3,4
1Experimental Ophthalmology, University of Geneva,1205 Geneva, Switzerland.
Abstract:
Oxidative stress (OS) is involved in the pathogenesis of retinal neurodegenerative diseases such as age-related macular degeneration (AMD) and diabetic retinopathy (DR) and an important target of therapeutic treatments. New therapeutics are tested in vivo despite limits in terms of transferability and ethical concerns. Retina cultures using human tissue can deliver critical information and significantly reduce the number of animal experiments along with increased transferability. We cultured up to 32 retina samples derived from one eye, analyzed the model's quality, induced OS, and tested the efficiency of antioxidative therapeutics. Bovine, porcine, rat, and human retinae were cultured in different experimental settings for 3-14 d. OS was induced by a high amount of glucose or hydrogen peroxide (H2O2) and treated with scutellarin, pigment epithelium-derived factor (PEDF), and/or granulocyte macrophage colony-stimulating factor (GM-CSF). The tissue morphology, cell viability, inflammation, and glutathione level were determined. The retina samples showed only moderate necrosis (23.83 ± 5.05 increased to 27.00 ± 1.66 AU PI-staining over 14 d) after 14 days in culture. OS was successfully induced (reduced ATP content of 288.3 ± 59.9 vs. 435.7 ± 166.8 nM ATP in the controls) and the antioxidants reduced OS-induced apoptosis (from 124.20 ± 51.09 to 60.80 ± 319.66 cells/image after the scutellarin treatment). Enhanced mammalian animal and human retina cultures enable reliable, highly transferable research on OS-triggered age-related diseases and pre-clinical testing during drug development.
Insights
Developing enhanced retinal cultures from human and animal tissues provides a reliable model for studying oxidative stress in diseases like AMD and DR. This method reduces animal use and improves therapeutic testing transferability.
Area of Science:
- Ophthalmology
- Neuroscience
- Toxicology
Background:
- Oxidative stress (OS) is a key factor in retinal neurodegenerative diseases, including age-related macular degeneration (AMD) and diabetic retinopathy (DR).
- Current in vivo testing of therapeutics faces limitations in transferability and ethical concerns regarding animal use.
Purpose of the Study:
- To establish and validate an enhanced mammalian and human retina culture model for studying OS-related retinal diseases.
- To assess the efficacy of antioxidative therapeutics in a controlled in vitro setting, reducing reliance on animal models.
Main Methods:
- Retinae from bovine, porcine, rat, and human sources were cultured for 3-14 days.
- Oxidative stress was induced using high glucose or hydrogen peroxide (H₂O₂).
- Treated samples with scutellarin, pigment epithelium-derived factor (PEDF), and granulocyte macrophage colony-stimulating factor (GM-CSF), assessing tissue morphology, cell viability, inflammation, and glutathione levels.
Main Results:
- Retina cultures demonstrated moderate necrosis (27.00 ± 1.66 AU PI-staining after 14 days).
- Successful induction of OS was confirmed by reduced ATP content (288.3 ± 59.9 nM vs. 435.7 ± 166.8 nM in controls).
- Antioxidants significantly reduced OS-induced apoptosis, with scutellarin treatment decreasing cell apoptosis.
Conclusions:
- Enhanced retina cultures offer a reliable and highly transferable model for OS research in age-related retinal diseases.
- This model facilitates pre-clinical testing of antioxidative therapeutics, aligning with ethical considerations and improving translational potential.
