Analysis of telomere lengths in human corneal endothelial cells from donors of different ages

C. A. Egan, I. Savre-Train, W. M. Boume, J. W. Shay

Research output: Contribution to journalArticle

Abstract

Purpose. To investigate the telornere hypothesis of cellular aging as the mechanism for cell cycle arrest in normal human corneal endothelium. Methods. The corneal endothelium and epithelium from human corneas from donors aged 6 weeks, 13 months, and 16, 31, 40, 54, and 84 years were dissected and frozen at -70°C. Purified DNA, digested with the restriction enzyme, Hinfl, was run on 0.7% agarose gels, probed with radiolabelled (AATCCC)4, and exposed to a Phosphorlmager screen (Molecular Dynamics). The length of the peak terminal restriction fragment (TRF) signal was determined by densitometry using ImageQuant software. Results. The endothelial cells had TRF lengths ranging from 10.8-12.0 kbp (mean 11.57). Corneal epithelial samples showed variable TRF lengths (mean 10.41. range 7.9-11.5). Conclusion. Human corneal endothelial cells have long telomeres. Their limited replicative ability is not explained by critically short telomere lengths.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number4
StatePublished - 1997

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Telomere
Corneal Endothelium
Endothelial Cells
Corneal Epithelium
Densitometry
Cell Aging
Molecular Dynamics Simulation
Cell Cycle Checkpoints
Sepharose
Cornea
Software
Gels
DNA
Enzymes

ASJC Scopus subject areas

  • Ophthalmology

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Analysis of telomere lengths in human corneal endothelial cells from donors of different ages. / Egan, C. A.; Savre-Train, I.; Boume, W. M.; Shay, J. W.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 4, 1997.

Research output: Contribution to journalArticle

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AU - Egan, C. A.

AU - Savre-Train, I.

AU - Boume, W. M.

AU - Shay, J. W.

PY - 1997

Y1 - 1997

N2 - Purpose. To investigate the telornere hypothesis of cellular aging as the mechanism for cell cycle arrest in normal human corneal endothelium. Methods. The corneal endothelium and epithelium from human corneas from donors aged 6 weeks, 13 months, and 16, 31, 40, 54, and 84 years were dissected and frozen at -70°C. Purified DNA, digested with the restriction enzyme, Hinfl, was run on 0.7% agarose gels, probed with radiolabelled (AATCCC)4, and exposed to a Phosphorlmager screen (Molecular Dynamics). The length of the peak terminal restriction fragment (TRF) signal was determined by densitometry using ImageQuant software. Results. The endothelial cells had TRF lengths ranging from 10.8-12.0 kbp (mean 11.57). Corneal epithelial samples showed variable TRF lengths (mean 10.41. range 7.9-11.5). Conclusion. Human corneal endothelial cells have long telomeres. Their limited replicative ability is not explained by critically short telomere lengths.

AB - Purpose. To investigate the telornere hypothesis of cellular aging as the mechanism for cell cycle arrest in normal human corneal endothelium. Methods. The corneal endothelium and epithelium from human corneas from donors aged 6 weeks, 13 months, and 16, 31, 40, 54, and 84 years were dissected and frozen at -70°C. Purified DNA, digested with the restriction enzyme, Hinfl, was run on 0.7% agarose gels, probed with radiolabelled (AATCCC)4, and exposed to a Phosphorlmager screen (Molecular Dynamics). The length of the peak terminal restriction fragment (TRF) signal was determined by densitometry using ImageQuant software. Results. The endothelial cells had TRF lengths ranging from 10.8-12.0 kbp (mean 11.57). Corneal epithelial samples showed variable TRF lengths (mean 10.41. range 7.9-11.5). Conclusion. Human corneal endothelial cells have long telomeres. Their limited replicative ability is not explained by critically short telomere lengths.

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