Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.

<h4>Background</h4>Cell viability and motility comprise ubiquitous mechanisms involved in a variety of (patho)biological processes including cancer. We report a technical comparative analysis of the novel impedance-based xCELLigence Real-Time Cell Analysis detection platform, with conven...

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Autores principales:	Ridha Limame, An Wouters, Bea Pauwels, Erik Fransen, Marc Peeters, Filip Lardon, Olivier De Wever, Patrick Pauwels
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/078390a688b14c2da2bad064c1ba0273
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spelling	oai:doaj.org-article:078390a688b14c2da2bad064c1ba02732021-11-18T08:11:31ZComparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.1932-620310.1371/journal.pone.0046536https://doaj.org/article/078390a688b14c2da2bad064c1ba02732012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23094027/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Cell viability and motility comprise ubiquitous mechanisms involved in a variety of (patho)biological processes including cancer. We report a technical comparative analysis of the novel impedance-based xCELLigence Real-Time Cell Analysis detection platform, with conventional label-based endpoint methods, hereby indicating performance characteristics and correlating dynamic observations of cell proliferation, cytotoxicity, migration and invasion on cancer cells in highly standardized experimental conditions.<h4>Methodology/principal findings</h4>Dynamic high-resolution assessments of proliferation, cytotoxicity and migration were performed using xCELLigence technology on the MDA-MB-231 (breast cancer) and A549 (lung cancer) cell lines. Proliferation kinetics were compared with the Sulforhodamine B (SRB) assay in a series of four cell concentrations, yielding fair to good correlations (Spearman's Rho 0.688 to 0.964). Cytotoxic action by paclitaxel (0-100 nM) correlated well with SRB (Rho>0.95) with similar IC(50) values. Reference cell migration experiments were performed using Transwell plates and correlated by pixel area calculation of crystal violet-stained membranes (Rho 0.90) and optical density (OD) measurement of extracted dye (Rho>0.95). Invasion was observed on MDA-MB-231 cells alone using Matrigel-coated Transwells as standard reference method and correlated by OD reading for two Matrigel densities (Rho>0.95). Variance component analysis revealed increased variances associated with impedance-based detection of migration and invasion, potentially caused by the sensitive nature of this method.<h4>Conclusions/significance</h4>The xCELLigence RTCA technology provides an accurate platform for non-invasive detection of cell viability and motility. The strong correlations with conventional methods imply a similar observation of cell behavior and interchangeability with other systems, illustrated by the highly correlating kinetic invasion profiles on different platforms applying only adapted matrix surface densities. The increased sensitivity however implies standardized experimental conditions to minimize technical-induced variance.Ridha LimameAn WoutersBea PauwelsErik FransenMarc PeetersFilip LardonOlivier De WeverPatrick PauwelsPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 10, p e46536 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ridha Limame An Wouters Bea Pauwels Erik Fransen Marc Peeters Filip Lardon Olivier De Wever Patrick Pauwels Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
description	<h4>Background</h4>Cell viability and motility comprise ubiquitous mechanisms involved in a variety of (patho)biological processes including cancer. We report a technical comparative analysis of the novel impedance-based xCELLigence Real-Time Cell Analysis detection platform, with conventional label-based endpoint methods, hereby indicating performance characteristics and correlating dynamic observations of cell proliferation, cytotoxicity, migration and invasion on cancer cells in highly standardized experimental conditions.<h4>Methodology/principal findings</h4>Dynamic high-resolution assessments of proliferation, cytotoxicity and migration were performed using xCELLigence technology on the MDA-MB-231 (breast cancer) and A549 (lung cancer) cell lines. Proliferation kinetics were compared with the Sulforhodamine B (SRB) assay in a series of four cell concentrations, yielding fair to good correlations (Spearman's Rho 0.688 to 0.964). Cytotoxic action by paclitaxel (0-100 nM) correlated well with SRB (Rho>0.95) with similar IC(50) values. Reference cell migration experiments were performed using Transwell plates and correlated by pixel area calculation of crystal violet-stained membranes (Rho 0.90) and optical density (OD) measurement of extracted dye (Rho>0.95). Invasion was observed on MDA-MB-231 cells alone using Matrigel-coated Transwells as standard reference method and correlated by OD reading for two Matrigel densities (Rho>0.95). Variance component analysis revealed increased variances associated with impedance-based detection of migration and invasion, potentially caused by the sensitive nature of this method.<h4>Conclusions/significance</h4>The xCELLigence RTCA technology provides an accurate platform for non-invasive detection of cell viability and motility. The strong correlations with conventional methods imply a similar observation of cell behavior and interchangeability with other systems, illustrated by the highly correlating kinetic invasion profiles on different platforms applying only adapted matrix surface densities. The increased sensitivity however implies standardized experimental conditions to minimize technical-induced variance.
format	article
author	Ridha Limame An Wouters Bea Pauwels Erik Fransen Marc Peeters Filip Lardon Olivier De Wever Patrick Pauwels
author_facet	Ridha Limame An Wouters Bea Pauwels Erik Fransen Marc Peeters Filip Lardon Olivier De Wever Patrick Pauwels
author_sort	Ridha Limame
title	Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
title_short	Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
title_full	Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
title_fullStr	Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
title_full_unstemmed	Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
title_sort	comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/078390a688b14c2da2bad064c1ba0273
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Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.

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