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A. P. Lynch, L. McGinley, C. Sheridan, C. Coleman, L. Howard, D. Kent, F. Barry; Investigating the Ability of Anti-Oxidant Gene Expression in Mesenchymal Stem Cells to Promote Cell Protection and Recovery in an in vitro Model of Age-Related Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1443.
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The aim of this study was to determine if lentiviral transduced mesenchymal stem cells (MSCs) expressing anti-oxidant genes could protect and recover the retinal pigment epithelial cell line, ARPE-19 from oxidative stress with tert-butylhydroperoxide (tert-BHP) treatment.
ARPE-19 cells were treated with varying concentrations of tert-BHP, ranging from 1 - 8 mM, to induce features of age-related macular degeneration (AMD). Control cells remained untreated. MSCs were transduced with lentiviral vectors expressing GFP, Catalase, HSP-27, HSP-70, SOD-1 or SOD-3. GFP transduced MSCs were used as a control. Treated ARPE-19 cells were grown in co-culture with transduced MSCs. Cell viability was assessed with the MTT and Live / Dead assays, while the production of ROS was measured with carboxy-H2DCFDA.
Treatment with tert-BHP decreased the viability of ARPE-19 cells. Morphological changes and cell survival assays revealed that incubation with transduced MSCs aided ARPE-19 cells to resist the free radicals induced by tert-BHP. In addition, activity of SOD-1 and -3 was enhanced in ARPE-19 cells leading to a reduction in ROS levels.
These results suggest that transduced MSCs have potent antioxidant activity and protect RPE from oxidative injury. Therefore transduced MSCs are possible candidates for the modulation of oxidative stress- induced damage of RPE in the ageing retina.
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