Abstract
Purpose: :
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.
Methods: :
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.
Results: :
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.
Conclusions: :
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.
Keywords: age-related macular degeneration • oxidation/oxidative or free radical damage • retinal pigment epithelium