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David R. Whikehart, Shilpa J. Register, Qing Chang, Brodrick Montgomery; Relationship of Telomeres and p53 in Aging Bovine Corneal Endothelial Cell Cultures. Invest. Ophthalmol. Vis. Sci. 2000;41(5):1070-1075.
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purpose. To demonstrate a relationship between telomere lengths and levels of
p53 in cultured bovine corneal endothelial cells (CECs) during aging.
methods. Bovine CECs were grown and aged as long-term cultures. Telomere lengths
were determined directly on gels with 32P probes after
treatment of isolated DNA with RsaI and HinfI. Protein p53 was determined using an enzyme-linked
immunosorbent sandwich assay. Cellular aging and the development of
replicative senescence were monitored by the appearance of senescent
morphology and the β-galactosidase assay.
results. Bovine CEC telomeres lost 4 kb (from 12.8 to 8.8 kb) over 1 year (89
population doublings [PDs]). The p53 levels in bovine CECs were
initially small (∼60 pg/million cells), but rose 3.5-fold by culture
age of 260 days (64 PDs). On initiation, cultured bovine CECs did not
stain for the senescent marker β-galactosidase. However, these cells
stained at 89 PDs and senescent morphology was observed in the cultures
at 64 PDs.
conclusions. The data indicate an inverse relationship between telomere lengths
(decreasing) and levels of p53 (increasing) in bovine CECs during
aging. These properties may influence the ability of these cells to
divide as they enter into replicative
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