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Shigeru Honda, Behnom Farboud, Leonard M. Hjelmeland, James T. Handa; Induction of an Aging mRNA Retinal Pigment Epithelial Cell Phenotype by Matrix-Containing Advanced Glycation End Products In Vitro. Invest. Ophthalmol. Vis. Sci. 2001;42(10):2419-2425.
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purpose. To determine an extensive mRNA phenotype of the established RPE cell
line ARPE-19 when grown on a matrix modified by advanced glycation end
methods. Growth Factor Reduced Matrigel (Collaborative Biomedical Products,
Bedford, MA) was nonenzymatically glycated with glycolaldehyde.
ARPE-19 cells were seeded on both AGE-Matrigel and Matrigel and grown
to confluence, and serum was withdrawn for 3 days. RNA was extracted,
and microarray analysis 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 cell
differentiation, growth factors that regulate the RPE cell and basement
membrane, and apoptosis. RT-PCR/Southern and Northern blot analysis
confirmed the expression patterns of selected genes, and flow cytometry
showed increased annexin V/propidium iodide-labeled cells when grown on
conclusions. Microarray analysis identified clusters of genes that could promote an
aging RPE phenotype in vitro induced by a matrix modified with
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