Aging of Human Fibroblast

Human fibroblast of different age were investigated for their gene expression profiles in quiescence. Cultures were established from skin samples derived from young and old donors, obtained from the NIA Aging Cell Repository (Coriell Institute for Medical Research, Camden, NJ). All cell lines originated from 2 mm punch biopsies taken from the medial aspect of the upper arm. The donors were members of the Baltimore Longitudinal Study of Aging (BLSA) where they were characterized as "healthy" indviduals. The cell lines investigated had normal karyotypes. RNA was isolated from the cell lysate using Qiagen RNeasy mini kit according to the manufacturer's instructions. Gene expression analysis was performed using the Codelink human bioarray containing single-stranded 30-mer oligonucleotide probes (Applied Microarrays, Tempe, AZ) and chips were run in duplicate. Details of this platform are available on the vendor's homepage website. Slides were scanned at 5 μm resolution with a ScanArray 4000 × l (Perkin Elmer, Waltham, Ma) and analyzed with the CodeLink Analysis Software, providing an integrated optical density (IOD) value for each hybridization spot, which is a measurement of an integrated background intensity value subtracted from the total pixel intensities within the area of the spot.