Rationale for the real-time and dynamic cell death assays using propidium iodide.

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dc.contributor.author Zhao, H en
dc.contributor.author Oczos, J en
dc.contributor.author Janowski, P en
dc.contributor.author Trembecka, D en
dc.contributor.author Dobrucki, J en
dc.contributor.author Darzynkiewicz, Z en
dc.contributor.author Wlodkowic, Donald en
dc.coverage.spatial United States en
dc.date.accessioned 2011-12-01T23:16:12Z en
dc.date.issued 2010-04 en
dc.identifier.citation Cytometry Part A 77A(4):399-405 Apr 2010 en
dc.identifier.issn 1552-4922 en
dc.identifier.uri http://hdl.handle.net/2292/9682 en
dc.description.abstract We have recently reported an innovative approach to use charged fluorochromes such as propidium iodide (PI) in the real-time, dynamic cell viability assays. This study was designed to provide a mechanistic rationale for the kinetic assays using cell permeability markers. Uptake of PI by live cells, effect on the cell cycle, long-term proliferation capacity, DNA damage response, and pharmacologic interactions with anticancer drugs were studied using both laser scanning microscopy and laser scanning cytometry. Exposure of human carcinomic alveolar basal epithelial A549 cells in cultures to 1.5 or 7.5 microM of PI for 24 h had minimal effect on cell cycle progression including DNA replication as measured by incorporation of 5'-ethynyl-2-deoxyuridine (EdU) detected by the "click chemistry" approach and measured by laser scanning cytometry. A modest reduction, from 44 to 40% or 33%, in frequency of DNA replicating cells was seen after 48 h at 1.5 or 7.5 microM concentration of PI. There was no evidence of increased phosphorylation of histone gammaH2AX in cells growing in the presence of 1.5 or 7.5 microM of PI for up to 48 h. Confocal image analysis of HeLa and NIH 3T3 mouse embryonic fibroblasts growing in the presence of PI showed granular distribution in cell cytoplasm suggesting PI accumulation in endosomes and progressive increase in fluorescence of nucleoli reflecting PI binding to nucleolar RNA. The overall responses of cells to cytotoxic agents were also not affected by the growth in the presence PI. Our data lend further support to the notion that PI can be effectively used in real-time, kinetic viability assays. en
dc.language eng en
dc.publisher Wiley-Blackwell en
dc.relation.ispartofseries Cytometry A en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. Details obtained from http://www.sherpa.ac.uk/romeo/issn/1552-4922/ en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.subject Biological Assay en
dc.subject Cell Death en
dc.subject Cell Line, Tumor en
dc.subject Cell Proliferation en
dc.subject DNA Replication en
dc.subject Fluorescence en
dc.subject Histones en
dc.subject Humans en
dc.subject Microscopy, Confocal en
dc.subject Phosphorylation en
dc.subject Propidium en
dc.subject Time Factors en
dc.title Rationale for the real-time and dynamic cell death assays using propidium iodide. en
dc.type Journal Article en
dc.identifier.doi 10.1002/cyto.a.20867 en
pubs.issue 4 en
pubs.begin-page 399 en
pubs.volume 77A en
dc.rights.holder Copyright: 2010 International Society for Advancement of Cytometry en
dc.identifier.pmid 20131407 en
pubs.end-page 405 en
dc.rights.accessrights http://purl.org/eprint/accessRights/RestrictedAccess en
pubs.subtype Article en
pubs.elements-id 230060 en
dc.identifier.eissn 1552-4930 en
pubs.record-created-at-source-date 2011-12-02 en
pubs.dimensions-id 20131407 en

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