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Impact of Storage Temperature on Gene Expression of Cultured ARPE ‐19 Cells
Author(s) -
Pasovic L.,
Eidet J.R.,
Olstad O.K.,
Chen D.F.,
Lyberg T.,
Utheim T.P.
Publication year - 2015
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2015.0481
Subject(s) - gene expression , gene , biology , dna microarray , microbiology and biotechnology , cell culture , cell , genetics
Purpose The development of a suitable storage method for retinal pigment epithelium ( RPE ) is necessary in the establishment of future RPE replacement therapy, and storage temperature has proven to be pivotal for cell survival. We analyzed the gene expression profile of cultured human RPE cells after storage at 4°C, 16°C, and 37°C. Methods ARPE ‐19 cells were cultured until confluence and stored in Minimum Essential Medium at 4°C, 16°C, and 37°C for seven days. DNA microarrays validated by qRT ‐ PCR were used to determine the gene expression profile. Results The gene expression profile of cultures stored at 4°C cluster closest to the control cultures that were not stored and display the least change in gene expression after storage (157 differentially expressed genes compared to controls), while cultures stored at 16°C and 37°C display much larger changes compared to controls (1787 and 1357 differentially expressed genes, respectively). Expression levels of key genes involved in phagocytosis, pigment synthesis, the visual cycle, cell junctions, and cellular transport were maintained close to control levels in cultures stored at both 4°C and 16°C in contrast to 37 °C. Conclusions RPE cell cultures stored at 16°C appear to modulate their gene expression profile in a manner that supports cell survival during storage, while maintaining the expression levels of genes important for key RPE functions.