Journal
OPHTHALMOLOGY AND THERAPY
Volume 3, Issue 1-2, Pages 49-61Publisher
SPRINGER INTERNATIONAL PUBLISHING AG
DOI: 10.1007/s40123-014-0024-9
Keywords
Additives; Age-related macular; degeneration; Ophthalmology; Retinal pigment epithelium; Storage; Viability
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Funding
- Norwegian Research Council
- SouthEastern Norway Regional Health Authority
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Introduction: There is increasing evidence that retinal pigment epithelium (RPE) can be used to treat age-related macular degeneration, one of the leading causes of blindness worldwide. However, the best way to store RPE to enable worldwide distribution is unknown. We investigated the effects of supplementing our previously published storage method with seven additives, attempting to improve the number of viable adult retinal pigment epithelial (ARPE)-19 cells after storage. Materials and methods: ARPE-19 cells were cultured on multiwell plates before being stored for 1 week at 16 degrees C. Unsupplemented Minimal Essential Medium (MEM) (control) and a total of seven individual additives (DADLE ([D-Ala 2, D-Leu 5]-encephalin), capsazepine, docosahexaenoic acid (DHA), resveratrol, quercetin, simvastatin and sulforaphane) at three to four concentrations in MEM were tested. The individual effect of each additive on cell viability was analyzed with a microplate fluorometer. Cell phenotype was investigated by both microplate fluorometer and epifluorescence microscopy, and morphology by scanning electron microscopy. Results: Supplementation of the storage medium with DADLE, capsazepine, DHA or resveratrol significantly increased the number of viable cells by 86.1% +/- 41.9%, 67.9% +/- 24.7%, 36.5% +/- 10.3% and 21.1% +/- 6.4%, respectively, compared to cells stored in unsupplemented MEM. DHA and resveratrol significantly reduced caspase-3 expression, while expression of RPE65 was maintained across groups. Conclusion: The number of viable ARPE-19 cells can be increased by the addition of DADLE, capsazepine, DHA or resveratrol to the storage medium without perturbing apoptosis or differentiation.
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