title: Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
abstract: BACKGROUND
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Center for Oncological Research (CORE), University of Antwerp, Antwerp, Belgium. ridha.limame@ua.ac.be
authors.forename: Ridha
authors.initials: R
authors.lastname: Limame
authors.forename: An
authors.initials: A
authors.lastname: Wouters
authors.forename: Bea
authors.initials: B
authors.lastname: Pauwels
authors.forename: Erik
authors.initials: E
authors.lastname: Fransen
authors.forename: Marc
authors.initials: M
authors.lastname: Peeters
authors.forename: Filip
authors.initials: F
authors.lastname: Lardon
authors.forename: Olivier
authors.initials: O
authors.lastname: De Wever
authors.forename: Patrick
authors.initials: P
authors.lastname: Pauwels
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D001681:Biological Assay; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen; D015194:Diffusion Chambers, Culture; D004338:Drug Combinations; D017097:Electric Impedance; D023381:Endpoint Determination; D005260:Female; D005840:Gentian Violet; D006801:Humans; D007797:Laminin; D017239:Paclitaxel / Q000494:pharmacology; D011509:Proteoglycans; D015203:Reproducibility of Results; D012235:Rhodamines; D012680:Sensitivity and Specificity
chemical_list: D000970:Antineoplastic Agents; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D012235:Rhodamines; C059485:matrigel; C022027:lissamine rhodamine B; D003094:Collagen; D005840:Gentian Violet; D017239:Paclitaxel