Using cell-substrate impedance and live cell imaging to measure real-time changes in cellular adhesion and de-adhesion induced by matrix modification.
Cell-matrix adhesion plays a key role in controlling cell morphology and signaling. Stimuli that disrupt cell-matrix adhesion (e.g., myeloperoxidase and other matrix-modifying oxidants/enzymes released during inflammation) are implicated in triggering pathological changes in cellular function, phenotype and viability in a number of diseases. Here, we describe how cell-substrate impedance and live cell imaging approaches can be readily employed to accurately quantify real-time changes in cell adhesion and de-adhesion induced by matrix modification (using endothelial cells and myeloperoxidase as a pathophysiological matrix-modifying stimulus) with high temporal resolution and in a non-invasive manner. The xCELLigence cell-substrate impedance system continuously quantifies the area of cell-matrix adhesion by measuring the electrical impedance at the cell-substrate interface in cells grown on gold microelectrode arrays. Image analysis of time-lapse differential interference contrast movies quantifies changes in the projected area of individual cells over time, representing changes in the area of cell-matrix contact. Both techniques accurately quantify rapid changes to cellular adhesion and de-adhesion processes. Cell-substrate impedance on microelectrode biosensor arrays provides a platform for robust, high-throughput measurements. Live cell imaging analyses provide additional detail regarding the nature and dynamics of the morphological changes quantified by cell-substrate impedance measurements. These complementary approaches provide valuable new insights into how myeloperoxidase-catalyzed oxidative modification of subcellular extracellular matrix components triggers rapid changes in cell adhesion, morphology and signaling in endothelial cells. These approaches are also applicable for studying cellular adhesion dynamics in response to other matrix-modifying stimuli and in related adherent cells (e.g., epithelial cells).
Centre for Vascular Research, University of New South Wales; m.rees@unsw.edu.au.
Martin D
MD
Rees
Centre for Vascular Research, University of New South Wales; School of Medical Sciences, University of New South Wales; shane.thomas@unsw.edu.au.
Shane R
SR
Thomas
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology*; D002478:Cells, Cultured; D017097:Electric Impedance; D042783:Endothelial Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D005109:Extracellular Matrix / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009195:Peroxidase / Q000737:chemistry / Q000378:metabolism
D016326:Extracellular Matrix Proteins; D009195:Peroxidase