1. Molecular Biology of Aging
2. These cell lines were initiated from primary explants taken antemortem from separate individuals and the patients are described as follows: age year:month; sex male (M) female (F); race caucasian (C); PDL population doubling (number of times cells divided to double the population); phenotype P NO NY or NM. Cell lines AG10548 (8:4 M C 5 P) AG10750 (9:4 M C 6 P) AG06297B (8:2 M C 33 P) AG12788 (90:00 M C 9 O) AG04064A (92:00 M C 15 O) AG04059B (96:00 M C 11 O) and AG11799 (37:00 M C 19 NM) were obtained from the Coriell Institute for Medical Research (Camden NJ). Cell lines CRL-1474 (7:00 M C 25 NY) CRL-7469 (9:00 M C 22 NY) and CRL-7581 (37:00 M C 20 NM) were obtained from the American Type Culture Collection (Manassas VA). Tables of data are available at Science Online at www.sciencemag.org/feature/data/1046164.shl. Cells were cultured (minimum essential medium Eagle–Earle balanced salt solution containing 2× concentrations of vitamins essential and nonessential amino acids and 10% fetal bovine serum) in 5% CO 2 at 37°C and were split in a 1:2 ratio at confluency with trypsin-EDTA. Bromodeoxyuridine (BrdU) incorporation was performed according to Becton Dickinson (Franklin Lakes NJ) protocols and imaged with epifluorescence microscopy. After a 48-hour incubation with 10 μM BrdU and staining with antibody to BrdU–fluorescein isothiocyanate (FITC) more than 90% of the fibroblasts derived from old-age and progeria individuals showed intense staining of the nucleus a similar ratio compared with cells from young and middle-age individuals. The analysis was based on 200 cells counted.
3. Progeria: a human-disease model of accelerated aging
4. Y Chromosome aneuploidy, micronuclei, kinetochores and aging in men
5. Cells were grown under identical conditions for three additional doublings to about 60% confluency. Polyadenylated RNAs were isolated and converted to cDNA followed by in vitro transcription cRNA synthesis. The biotinylated cRNAs were then hybridized to the HuGeneFL oligonucleotide arrays (Affymetrix Santa Clara CA) which contain probes for more than 6000 known human genes. All procedures were performed according to prescribed Affymetrix protocols.