title: Effect of Storage Temperature on Key Functions of Cultured Retinal Pigment Epithelial Cells.
abstract: Purpose. Replacement of the diseased retinal pigment epithelium (RPE) with cells capable of performing the specialized functions of the RPE is the aim of cell replacement therapy for treatment of macular degenerative diseases. A storage method for RPE is likely to become a prerequisite for the establishment of such treatment. Herein, we analyze the effect of storage temperature on key functions of cultured RPE cells. Methods. Cultured ARPE-19 cells were stored in Minimum Essential Medium at 4°C, 16°C, and 37°C for seven days. Total RNA was isolated and the gene expression profile was determined using DNA microarrays. Comparison of the microarray expression values with qRT-PCR analysis of selected genes validated the results. Results. Expression levels of several key genes involved in phagocytosis, pigment synthesis, the visual cycle, adherens, and tight junctions, and glucose and ion transport were maintained close to control levels in cultures stored at 4°C and 16°C. Cultures stored at 37°C displayed regulational changes in a larger subset of genes related to phagocytosis, adherens, and tight junctions. Conclusion. RPE cultures stored at 4°C and 16°C for one week are capable of maintaining the expression levels of genes important for key RPE functions close to control levels.
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway ; Faculty of Medicine, University of Oslo, Sognsvannsveien 9, 0372 Oslo, Norway.
authors.forename: Lara
authors.initials: L
authors.lastname: Pasovic
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway.
authors.forename: Jon Roger
authors.initials: JR
authors.lastname: Eidet
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway.
authors.forename: Berit S
authors.initials: BS
authors.lastname: Brusletto
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway.
authors.forename: Torstein
authors.initials: T
authors.lastname: Lyberg
authors.affiliation: Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway ; Department of Oral Biology, Faculty of Dentistry, University of Oslo, Sognsvannsveien 10, P.O. Box 1052, Blindern, 0316 Oslo, Norway.
authors.forename: Tor P
authors.initials: TP
authors.lastname: Utheim