title: Antioxidants Improve the Viability of Stored Adult Retinal Pigment Epithelial-19 Cultures.
abstract: BACKGROUND
  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.
  
  
  METHODS
  ARPE-19 cells were cultured on multiwell plates before being stored for 1 week at 16 °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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway. larapasovic@gmail.com.
authors.forename: Lara
authors.initials: L
authors.lastname: Pasovic
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.
authors.forename: Jon R
authors.initials: JR
authors.lastname: Eidet
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.
authors.forename: Torstein
authors.initials: T
authors.lastname: Lyberg
authors.affiliation: Department of Oral Biology, University of Oslo, Oslo, Norway.
authors.forename: Edward B
authors.initials: EB
authors.lastname: Messelt
authors.affiliation: Division of Surgery, Ear, Nose and Throat Department, Akershus University Hospital, Lørenskog, Norway.
authors.forename: Peder
authors.initials: P
authors.lastname: Aabel
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.
authors.forename: Tor P
authors.initials: TP
authors.lastname: Utheim
keywords: Additives; Age-related macular degeneration; Ophthalmology; Retinal pigment epithelium; Storage; Viability