title: Electric cell-substrate impedance sensing for the quantification of endothelial proliferation, barrier function, and motility.
abstract: Electric Cell-substrate Impedance Sensing (ECIS) is an in vitro impedance measuring system to quantify the behavior of cells within adherent cell layers. To this end, cells are grown in special culture chambers on top of opposing, circular gold electrodes. A constant small alternating current is applied between the electrodes and the potential across is measured. The insulating properties of the cell membrane create a resistance towards the electrical current flow resulting in an increased electrical potential between the electrodes. Measuring cellular impedance in this manner allows the automated study of cell attachment, growth, morphology, function, and motility. Although the ECIS measurement itself is straightforward and easy to learn, the underlying theory is complex and selection of the right settings and correct analysis and interpretation of the data is not self-evident. Yet, a clear protocol describing the individual steps from the experimental design to preparation, realization, and analysis of the experiment is not available. In this article the basic measurement principle as well as possible applications, experimental considerations, advantages and limitations of the ECIS system are discussed. A guide is provided for the study of cell attachment, spreading and proliferation; quantification of cell behavior in a confluent layer, with regard to barrier function, cell motility, quality of cell-cell and cell-substrate adhesions; and quantification of wound healing and cellular responses to vasoactive stimuli. Representative results are discussed based on human microvascular (MVEC) and human umbilical vein endothelial cells (HUVEC), but are applicable to all adherent growing cells.
authors.affiliation: Department of Pulmonary Diseases, Institute for Cardiovascular Research, VU University Medical Center.
authors.forename: Robert
authors.initials: R
authors.lastname: Szulcek
authors.affiliation: Department of Pulmonary Diseases, Institute for Cardiovascular Research, VU University Medical Center.
authors.forename: Harm Jan
authors.initials: HJ
authors.lastname: Bogaard
authors.affiliation: Department of Physiology, Institute for Cardiovascular Research, VU University Medical Center; nieuwamerongen@vumc.nl.
authors.forename: Geerten P
authors.initials: GP
authors.lastname: van Nieuw Amerongen
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication / Q000502:physiology*; D048708:Cell Growth Processes / Q000502:physiology; D002465:Cell Movement / Q000502:physiology*; D017097:Electric Impedance; D042783:Endothelial Cells / Q000166:cytology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D055806:Microvessels / Q000166:cytology / Q000502:physiology
references.citation: Circ Res. 2000 Aug 18;87(4):335-40
references.pmid: 10948069
references.citation: Adv Biochem Eng Biotechnol. 2008;109:195-237
references.pmid: 17992488
references.citation: Pflugers Arch. 1999 May;437(6):925-34
references.pmid: 10370072
references.citation: Ann Biomed Eng. 2001 Aug;29(8):648-56
references.pmid: 11556721
references.citation: Am J Physiol Cell Physiol. 2011 Jan;300(1):C146-54
references.pmid: 20861463
references.citation: Cardiovasc Res. 2013 Aug 1;99(3):471-82
references.pmid: 23536606
references.citation: Exp Cell Res. 1993 Jan;204(1):102-9
references.pmid: 8416788
references.citation: Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7896-900
references.pmid: 1881923
references.citation: Nat Protoc. 2007;2(2):329-33
references.pmid: 17406593
references.citation: J Biochem Biophys Methods. 1996 Jul 10;32(3):151-70
references.pmid: 8844323
references.citation: Circulation. 2012 Dec 4;126(23):2728-38
references.pmid: 23099479
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 10;101(6):1554-9
references.pmid: 14747654
references.citation: PLoS One. 2013;8(3):e57423
references.pmid: 23483910
references.citation: Nature. 1993 Dec 9;366(6455):591-2
references.pmid: 8255299
references.citation: Biophys J. 1995 Dec;69(6):2800-7
references.pmid: 8599686
references.citation: Exp Cell Res. 2000 Aug 25;259(1):158-66
references.pmid: 10942588
references.citation: Exp Cell Res. 1999 Aug 1;250(2):576-80
references.pmid: 10413610
references.citation: J Vis Exp. 2012 Apr 01;(62):
references.pmid: 22491349
references.citation: Biosens Bioelectron. 2009 Apr 15;24(8):2625-9
references.pmid: 19230649