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Journal of Ophthalmology
Volume 2013 (2013), Article ID 216359, 11 pages
http://dx.doi.org/10.1155/2013/216359
Research Article

Optimization of Storage Temperature for Cultured ARPE-19 Cells

1Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway
2Department of Ophthalmology, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway
3SynsLaser Kirurgi Oslo/Tromsø, Lille Grensen 7, 0159 Oslo, Norway
4Department of Oral Biology, Faculty of Dentistry, University of Oslo, Sognsvannsveien 10, P.O. Box 1052, Blindern, 0316 Oslo, Norway
5Division of Surgery, Ear, Nose and Throat Department, Akershus University Hospital, Sykehusveien 25, 1478 Lørenskog, Norway
6Schepens Eye Research Institute, Department of Ophthalmology, Harvard Medical School, 20 Staniford Street, Boston, MA 02114, USA

Received 4 July 2013; Revised 2 September 2013; Accepted 3 September 2013

Academic Editor: Alfredo García-Layana

Copyright © 2013 Lara Pasovic et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose. The establishment of future retinal pigment epithelium (RPE) replacement therapy is partly dependent on the availability of tissue-engineered RPE cells, which may be enhanced by the development of suitable storage methods for RPE. This study investigates the effect of different storage temperatures on the viability, morphology, and phenotype of cultured RPE. Methods. ARPE-19 cells were cultured under standard conditions and stored in HEPES-buffered MEM at nine temperatures (4°C, 8°C, 12°C, 16°C, 20°C, 24°C, 28°C, 32°C, and 37°C) for seven days. Viability and phenotype were assessed by a microplate fluorometer and epifluorescence microscopy, while morphology was analyzed by scanning electron microscopy. Results. The percentage of viable cells preserved after storage was highest in the 16°C group ( %; compared to 4°C, 8°C, and 24°C–37°C; compared to 12°C). Ultrastructure was best preserved at 12°C, 16°C, and 20°C. Expression of actin, ZO-1, PCNA, caspase-3, and RPE65 was maintained after storage at 16°C compared to control cells that were not stored. Conclusion. Out of nine temperatures tested between 4°C and 37°C, storage at 12°C, 16°C, and 20°C was optimal for maintenance of RPE cell viability, morphology, and phenotype. The preservation of RPE cells is critically dependent on storage temperature.