Induction of an aging mRNA retinal pigment epithelial cell phenotype by matrix-containing advanced glycation end products in vitro

PURPOSE. To determine an extensive mRNA phenotype of the established RPE ce ll line ARPE-19 when grown on a matrix modified by advanced glycation end p roducts (AGES). METHODS. Growth Factor Reduced Matrigel (Collaborative Biomedical Products, Bedford, MA) was nonenzymatically glycated with glycolaldehyde. ARPE-19 ce lls were seeded on both AGE-Matrigel and Matrigel and grown to confluence, and serum was withdrawn for 3 days. RNA was extracted, and microarray analy sis was performed to characterize the genes, which are altered by a matrix modified by AGES. Gene expression changes were confirmed by RT-PCR/Southern and Northern blot analysis. Apoptosis was measured by annexin V/propidium iodide labeling. RESULTS. Clusters of genes with altered expression were found related to ce ll differentiation, growth factors that regulate the RPE cell and basement membrane, and apoptosis. RT-PCR/Southern and Not-them blot analysis confirm ed the expression patterns of selected genes, and flow cytometry showed inc reased annexin V/propidium iodide-labeled cells when grown on AGE-Matrigel. CONCLUSIONS. Microarray analysis identified clusters of genes that could pr omote an aging RPE phenotype in vitro induced by a matrix modified with AGE S.