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  • Senescence associated β galactosidase activity in human retinal pigment epithelial cells exposed to mild hyperoxia in vitro

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Scientific correspondence
Senescence associated β galactosidase activity in human retinal pigment epithelial cells exposed to mild hyperoxia in vitro
  1. Shigeru Honda1,
  2. Leonard M Hjelmeland2,
  3. James T Handa1
  1. 1Department of Ophthalmology, University of California, Davis, CA, USA
  2. 2Departments of Ophthalmology and Molecular and Cellular Biology University of California, Davis, CA, USA
  1. Correspondence to: James T Handa, MD, 3-109 Jefferson Building, Johns Hopkins Hospital, 600 N Wolfe Street, Baltimore, MD 21287, USA; jthanda{at}jhmi.edu

Abstract

Aims: To determine if mild hyperoxia induces senescence in retinal pigment epithelium (RPE) cells in vitro.

Methods: RPE340 cells and WI38 cells were grown in 20% oxygen and 40% oxygen until proliferative exhaustion. A combined senescence associated β galactosidase (SABG) and 5-bromo-2`-deoxyuridine (BrdU) double labelling technique was performed at different times and labelled cells were counted.

Results: Cells grown in 40% oxygen stopped proliferating at an earlier population doubling level than when grown in 20% oxygen. An increase in SABG positive cells and decrease of BrdU positive cells in 40% oxygen developed at an earlier time than when grown in 20% oxygen.

Conclusion: Mild hyperoxia induces premature senescence in a specific RPE cell line.

  • BrdU, 5-bromo-2`-deoxyuridine
  • PDL, population doubling level
  • RPE, retinal pigment epithelium
  • SABG, senescent associated β galactosidase

https://doi.org/10.1136/bjo.86.2.159

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