These authors contributed equally to this work.
Characterization of cellular senescence mechanisms in human corneal endothelial cells
Article first published online: 29 DEC 2011
© 2011 The Authors. Aging Cell © 2011 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland
Volume 11, Issue 2, pages 234–240, April 2012
How to Cite
Sheerin, A. N., Smith, S. K., Jennert-Burston, K., Brook, A. J., Allen, M. C., Ibrahim, B., Jones, D., Wallis, C., Engelmann, K., Rhys-Williams, W., Faragher, R. G. A. and Kipling, D. (2012), Characterization of cellular senescence mechanisms in human corneal endothelial cells. Aging Cell, 11: 234–240. doi: 10.1111/j.1474-9726.2011.00776.x
Re-use of this article is permitted in accordance with the Terms and Conditions set out at http://wileyonlinelibrary.com/onlineopen#OnlineOpen_Terms
- Issue published online: 15 MAR 2012
- Article first published online: 29 DEC 2011
- Accepted manuscript online: 29 NOV 2011 10:47AM EST
- Accepted for publication 21 November 2011
- oxidative stress;
- replicative senescence
The human cornea is a tri-laminar structure composed of several cell types with substantial mitotic potential. Age-related changes in the cornea are associated with declining visual acuity and the onset of overt age-related corneal diseases. Corneal transplantation is commonly used to restore vision in patients with damaged or diseased corneas. However, the supply of donor tissue is limited, and thus there is considerable interest in the development of tissue-engineered alternatives. A major obstacle to these approaches is the short replicative lifespan of primary human corneal endothelial cells (HCEC). Accordingly, a comprehensive investigation of the signalling pathways and mechanisms underpinning proliferative lifespan and senescence in HCEC was undertaken. The effects of exogenous human telomerase reverse transcriptase expression, p53 knockdown, disruption of the pRb pathway by over-expression of CDK4 and reduced oxygen concentration on the lifespan of primary HCEC were evaluated. We provide proof-of-principle that forced expression of telomerase, when combined with either p53 knockdown or CDK4 over-expression, is sufficient to produce immortalized HCEC lines. The resultant cell lines express an HCEC-specific transcriptional fingerprint, and retain expression of the corneal endothelial temperature-sensitive potassium channel, suggesting that significant dedifferentiation does not occur as a result of these modes of immortalization. Exploiting these insights into proliferative lifespan barriers in HCEC will underpin the development of novel strategies for cell-based therapies in the human cornea.