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

Autor: Bea Pauwels, Marc Peeters, Ridha Limame, Filip Lardon, An Wouters, Erik Fransen, Olivier De Wever, Patrick Pauwels
Rok vydání: 2012
Předmět:
Cell
Sulforhodamine B
Cytopathology
chemistry.chemical_compound
Cell Movement
Molecular Cell Biology
Basic Cancer Research
Electric Impedance
Morphogenesis
Pathology
Cytotoxicity
Multidisciplinary
Cell migration
Cell Differentiation
CANCER
Drug Combinations
Cell Motility
medicine.anatomical_structure
Oncology
Diffusion Chambers
Culture
Medicine
Biological Assay
Female
Proteoglycans
Collagen
Engineering sciences. Technology
Research Article
Test Evaluation
Paclitaxel
Cell Survival
Endpoint Determination
Science
Biophysics
Antineoplastic Agents
Cell Migration
Biology
Sensitivity and Specificity
Cell Growth
Diagnostic Medicine
Cell Line
Tumor
medicine
Humans
Viability assay
Cell Proliferation
Matrigel
Cell growth
Rhodamines
Biology and Life Sciences
Reproducibility of Results
Cancers and Neoplasms
IN-VITRO
chemistry
Anatomical Pathology
Immunology
Cancer cell
Gentian Violet
Laminin
Developmental Biology
Zdroj: PLoS ONE
PLoS ONE, Vol 7, Iss 10, p e46536 (2012)
PLOS ONE
ISSN: 1932-6203
Popis: BackgroundCell 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.Methodology/principal findingsDynamic 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.Conclusions/significanceThe 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.
Databáze: OpenAIRE
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