Assessment of oxidative damage in the primary mouse ocular surface cells/stem cells in response to ultraviolet-c (UV-C) damage

Bipasha Bose, Saketh Kapoor, Utsav Sen, A. S. Muhammad Nihad, Debajit Chaudhury, P. Sudheer Shenoy

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

The ocular surface is subjected to regular wear and tear due to various environmental factors. Exposure to UV-C radiation constitutes an occupational health hazard. Here, we demonstrate the exposure of primary stem cells from the mouse ocular surface to UV-C radiation. Reactive oxygen species (ROS) formation is the readout of the extent of oxidative stress/damage. In an experimental in vitro setting, it is also essential to assess the percentage of dead cells generated due to oxidative stress. In this article, we will demonstrate the 2',7'-Dichlorofluoresceindiacetate (DCFDA) staining of UV-C exposed mouse primary ocular surface stem cells and their quantification based on the fluorescent images of DCFDA staining. DCFDA staining directly corresponds to ROS generation. We also demonstrate the quantification of dead and live cells by simultaneous staining with propidium iodide (PI) and Hoechst 3332 respectively and the percentage of DCFDA (ROS positive) and PI positive cells.

Original languageEnglish
Article numbere59924
JournalJournal of Visualized Experiments
Volume2020
Issue number156
DOIs
StatePublished - Feb 2020
Externally publishedYes

Keywords

  • 2'
  • 7'-dichlorofluoresceindiacetate (DCFDA)
  • Developmental Biology
  • Issue 156
  • Live/dead cell detection
  • Primary ocular stem cells
  • Reactive oxygen species (ROS)
  • UV-C
  • UV-C damage

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