Assessment of cytotoxicity by impedance spectroscopy.

This paper describes a simple and convenient method to monitor on-line cell adhesion by electrical impedance measurements. Immortalized mouse fibroblasts, BALB/3T3, were cultured onto interdigitated electrode structures integrated into the bottom of an in-house fabricated device. Impedance modulus, phase, real and imaginary parts were considered separately and plotted as function of frequency and time to better understand and select the component giving more information on cell adhesion changes. For cytotoxicity assessment, the cells were treated with different concentrations of sodium arsenite used as model toxicant and their responses were monitored on-line. The half inhibition concentration, the required concentration to achieve 50% inhibition, derived from the measurements fall between the results obtained using standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test and colony forming efficiency assay confirming the good sensitivity of the system. In term of impedance signal, the modulus results was found to be the most sensitive of the considered components for cytotoxicity testing of chemicals.

[1]  John H T Luong,et al.  Assessment of cytotoxicity by emerging impedance spectroscopy. , 2005, Toxicology and applied pharmacology.

[2]  Bernard Lachance,et al.  Assessment of cytotoxicity using electric cell-substrate impedance sensing: concentration and time response function approach. , 2002, Analytical chemistry.

[3]  Joachim Wegener,et al.  Use of electrochemical impedance measurements to monitor β-adrenergic stimulation of bovine aortic endothelial cells , 1999, Pflügers Archiv.

[4]  R. McGuinness,et al.  Cellular Dielectric Spectroscopy: A Powerful New Approach to Label-Free Cellular Analysis , 2004, Journal of biomolecular screening.

[5]  Bernard Lachance,et al.  An in-depth analysis of electric cell-substrate impedance sensing to study the attachment and spreading of mammalian cells. , 2002, Analytical chemistry.

[6]  Joachim Wegener,et al.  Bioelectrical impedance assay to monitor changes in cell shape during apoptosis. , 2004, Biosensors & bioelectronics.

[7]  I. Giaever,et al.  Monitoring fibroblast behavior in tissue culture with an applied electric field. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[8]  E. Testai,et al.  Toxicology investigations with cell culture systems: 20 years after. , 2004, Toxicology in vitro : an international journal published in association with BIBRA.

[9]  John H T Luong,et al.  On‐Line Monitoring of Cell Growth and Cytotoxicity Using Electric Cell‐Substrate Impedance Sensing (ECIS) , 2003, Biotechnology progress.

[10]  Wei Zheng,et al.  Application of real-time cell electronic sensing (RT-CES) technology to cell-based assays. , 2004, Assay and drug development technologies.

[11]  Xiao Xu,et al.  Real-time monitoring of morphological changes in living cells by electronic cell sensor arrays: an approach to study G protein-coupled receptors. , 2006, Analytical chemistry.

[12]  Ralf Ehret,et al.  Real-time assessment of cytotoxicity by impedance measurement on a 96-well plate , 2007 .

[13]  Ralf Ehret,et al.  Microelectronic sensor system for microphysiological application on living cells , 1999 .

[14]  H. Galla,et al.  Impedance analysis of epithelial and endothelial cell monolayers cultured on gold surfaces. , 1996, Journal of biochemical and biophysical methods.

[15]  B Wolf,et al.  Approach to a multiparametric sensor-chip-based tumor chemosensitivity assay , 2001, Anti-cancer drugs.

[16]  Stephan Gabos,et al.  Dynamic monitoring of cytotoxicity on microelectronic sensors. , 2005, Chemical research in toxicology.

[17]  H Babich,et al.  Comparisons of two in vitro cytotoxicity assays-The neutral red (NR) and tetrazolium MTT tests. , 1988, Toxicology in vitro : an international journal published in association with BIBRA.

[18]  Fibroblast growth and H‐7 protein kinase inhibitor response monitored in microimpedance sensor arrays , 2004, Biotechnology and bioengineering.

[19]  B Wolf,et al.  Monitoring of cellular behaviour by impedance measurements on interdigitated electrode structures. , 1997, Biosensors & bioelectronics.

[20]  Ju Hun Yeon,et al.  Cytotoxicity test based on electrochemical impedance measurement of HepG2 cultured in microfabricated cell chip. , 2005, Analytical biochemistry.

[21]  J. Wegener,et al.  Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces. , 2000, Experimental cell research.

[22]  T. Hug,et al.  Biophysical methods for monitoring cell-substrate interactions in drug discovery. , 2003, Assay and drug development technologies.

[23]  Jenny Zhu,et al.  Label-free, real-time monitoring of IgE-mediated mast cell activation on microelectronic cell sensor arrays. , 2004, Journal of immunological methods.