2.50
Hdl Handle:
http://hdl.handle.net/10755/157489
Type:
Presentation
Title:
PLATE-BASED ASSAYS TO MEASURE REACTIVE OXYGEN SPECIES IN MODELS OF AGING
Abstract:
PLATE-BASED ASSAYS TO MEASURE REACTIVE OXYGEN SPECIES IN MODELS OF AGING
Conference Sponsor:Western Institute of Nursing
Conference Year:2010
Author:Downs, Charles A., MSN, ACNP-BC
P.I. Institution Name:University of Arizona
Title:Doctoral Student
Contact Address:1305 N. Martin, PO Box 210203, Tucson, AZ, 85721, USA
Co-Authors:Carrie J. Merkle; David W. Montgomery
PURPOSE: Our laboratory uses cell culture models to study aging-related phenomena. It is becoming increasing apparent that reactive oxygen species (ROS) have a role in aging and age-related diseases. The purpose of this paper is to describe plate-based assays in our laboratory to assess ROS in cell culture models of aging.
BACKGROUND: Excessive ROS production contributes to biological aging and age-related diseases such as atherosclerosis and cancer. Furthermore, ROS can contribute to cancer growth and metastases. We have used plate-based assays to study intracellular and extracellular ROS production by aged vascular endothelial cells after breast cancer cell addition. Intracellular ROS is measured using the membrane-permeant probe, dichlorodihydrofluorescein diacetate (CM-H2DCFDA), which is converted to an impermeant form by intracellular esterases. In the presence of ROS (e.g. nitric oxide, superoxide, peroxide) the probe fluoresces and the fluorescence can be quantified using a plate reader. Extracellular hydrogen peroxide is measured using the Amplex Red reagent (dihydroxyphenoxazine) that in combination with horseradish peroxidase reacts with hydrogen peroxide to produce a measureable fluorescent product. These assays permit assessment of intracellular or extracellular ROS production in response to various treatments or changes in physiologic conditions.
METHODS: We have used these plate-based assays to determine why old and young bovine pulmonary artery endothelial cells (BPAECs) respond differently to breast cancer cell addition. We found, to our surprise, that it was due to an increase production of ROS by the BPAECs in contrast to ROS production by cancer cells reported in the literature. Currently these assays are being used to understand age-related pulmonary disease using cell culture models of alveolar type I and microvascular endothelial cells.
RESULTS:/IMPLICATIONS: These assays have been useful in measuring intracellular and extracellular ROS production in a variety of cell types and models. This will increase our understanding of ROS in aging and age-related diseases and may assist in the development of nursing interventions to ameliorate the effects of ROS.
Repository Posting Date:
26-Oct-2011
Date of Publication:
17-Oct-2011
Sponsors:
Western Institute of Nursing

Full metadata record

DC FieldValue Language
dc.typePresentationen_GB
dc.titlePLATE-BASED ASSAYS TO MEASURE REACTIVE OXYGEN SPECIES IN MODELS OF AGINGen_GB
dc.identifier.urihttp://hdl.handle.net/10755/157489-
dc.description.abstract<table><tr><td colspan="2" class="item-title">PLATE-BASED ASSAYS TO MEASURE REACTIVE OXYGEN SPECIES IN MODELS OF AGING</td></tr><tr class="item-sponsor"><td class="label">Conference Sponsor:</td><td class="value">Western Institute of Nursing</td></tr><tr class="item-year"><td class="label">Conference Year:</td><td class="value">2010</td></tr><tr class="item-author"><td class="label">Author:</td><td class="value">Downs, Charles A., MSN, ACNP-BC</td></tr><tr class="item-institute"><td class="label">P.I. Institution Name:</td><td class="value">University of Arizona</td></tr><tr class="item-author-title"><td class="label">Title:</td><td class="value">Doctoral Student</td></tr><tr class="item-address"><td class="label">Contact Address:</td><td class="value">1305 N. Martin, PO Box 210203, Tucson, AZ, 85721, USA</td></tr><tr class="item-email"><td class="label">Email:</td><td class="value">cdowns@nursing.arizona.edu</td></tr><tr class="item-co-authors"><td class="label">Co-Authors:</td><td class="value">Carrie J. Merkle; David W. Montgomery</td></tr><tr><td colspan="2" class="item-abstract">PURPOSE: Our laboratory uses cell culture models to study aging-related phenomena. It is becoming increasing apparent that reactive oxygen species (ROS) have a role in aging and age-related diseases. The purpose of this paper is to describe plate-based assays in our laboratory to assess ROS in cell culture models of aging. <br/>BACKGROUND: Excessive ROS production contributes to biological aging and age-related diseases such as atherosclerosis and cancer. Furthermore, ROS can contribute to cancer growth and metastases. We have used plate-based assays to study intracellular and extracellular ROS production by aged vascular endothelial cells after breast cancer cell addition. Intracellular ROS is measured using the membrane-permeant probe, dichlorodihydrofluorescein diacetate (CM-H2DCFDA), which is converted to an impermeant form by intracellular esterases. In the presence of ROS (e.g. nitric oxide, superoxide, peroxide) the probe fluoresces and the fluorescence can be quantified using a plate reader. Extracellular hydrogen peroxide is measured using the Amplex Red reagent (dihydroxyphenoxazine) that in combination with horseradish peroxidase reacts with hydrogen peroxide to produce a measureable fluorescent product. These assays permit assessment of intracellular or extracellular ROS production in response to various treatments or changes in physiologic conditions. <br/>METHODS: We have used these plate-based assays to determine why old and young bovine pulmonary artery endothelial cells (BPAECs) respond differently to breast cancer cell addition. We found, to our surprise, that it was due to an increase production of ROS by the BPAECs in contrast to ROS production by cancer cells reported in the literature. Currently these assays are being used to understand age-related pulmonary disease using cell culture models of alveolar type I and microvascular endothelial cells. <br/>RESULTS:/IMPLICATIONS: These assays have been useful in measuring intracellular and extracellular ROS production in a variety of cell types and models. This will increase our understanding of ROS in aging and age-related diseases and may assist in the development of nursing interventions to ameliorate the effects of ROS. <br/></td></tr></table>en_GB
dc.date.available2011-10-26T19:55:07Z-
dc.date.issued2011-10-17en_GB
dc.date.accessioned2011-10-26T19:55:07Z-
dc.description.sponsorshipWestern Institute of Nursingen_GB
All Items in this repository are protected by copyright, with all rights reserved, unless otherwise indicated.