Publication Cover
Endothelium
Journal of Endothelial Cell Research
Volume 6, 1998 - Issue 2
16
Views
12
CrossRef citations to date
0
Altmetric
Original Article

A Sensitive Fluorometric Assay for Determining Hydrogen Peroxide-mediated Sublethal and Lethal Endothelial Cell Injury

, &
Pages 143-151 | Received 05 Feb 1998, Published online: 13 Jul 2009
 

Abstract

A rapid and sensitive quantitative fluorometric assay was developed to measure the response of endothelial cells to hydrogen peroxide (H2O2). The response of an endothelial cell-derived cell line, EA-hy-926, or human umbilical vein endothelial cells to H2O2 was determined using calcein-AM, a dye which becomes fluorescent upon cleavage by intracellular esterase(s). The ability of the cells to take up and convert calcein-AM was measured directly in the wells of 96-well flat-bottomed tissue culture plates with cell monolayers using a computerised microplate fluorimeter, or in cell suspensions using flow cytometry. The results obtained by these techniques were compared with each other and with a standard 51Cr release cytotoxicity assay. We found that calcein-AM is a highly sensitive probe for measuring H2O2-mediated cell injury, as it can not only detect the irreversible cyotoxicity measured by 51Cr release assay, but can also distinguish sublethal and reversible injury seen at low H2O2 concentrations.

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.