Cytotoxicity test based on electrochemical impedance measurement of HepG2 cultured in microfabricated cell chip.

This paper presents the use of electrochemical impedance measurement on a cell chip to monitor cell growth as a consequence of treatment with potentially cytotoxic agents. The cell chip consists of an eight-well cell culture chamber incorporated with a three-electrode system on each well. The gold electrode for impedance measurements is fabricated by sputtering on polycarbonate film. Human hepatocellular carcinoma cell (HepG2) is adapted to cytotoxicity test using the cell chip. Although the relatively small quantity of cells on the electrode has been measured indirectly, the cell chip can monitor toxic effects on the HepG2 cells cultured in the cell chip continuously and detect cellular behavior without multiple reagents. The cells in the stationary phase after plating are used for the cytotoxicity experiment and the impedance is decreased after treatments with several toxicants, such as tamoxifen and menadione, indicating the detachment of dead cells. These results reveal that the microfabricated cell chip system provides an easy and real-time monitoring method for cytotoxicity test.

[1]  Allen J. Bard,et al.  Electrochemical Methods: Fundamentals and Applications , 1980 .

[2]  J. Teissié,et al.  Chinese hamster ovary cells sensitivity to localized electrical stresses. , 1999, Bioelectrochemistry and bioenergetics.

[3]  C. Lieber,et al.  Toxicity of β-carotene and its exacerbation by acetaldehyde in HepG2 cells , 2001 .

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

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

[6]  Yoo,et al.  An electrochemical impedance measurement technique employing Fourier transform , 2000, Analytical chemistry.

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

[8]  C. Mothersill,et al.  In vitro cytotoxicity testing of three zinc metal salts using established fish cell lines. , 2004, Toxicology in vitro : an international journal published in association with BIBRA.

[9]  A. Li,et al.  Cryopreserved human hepatocytes: characterization of drug-metabolizing enzyme activities and applications in higher throughput screening assays for hepatotoxicity, metabolic stability, and drug-drug interaction potential. , 1999, Chemico-biological interactions.

[10]  N. Depaola,et al.  Spatial variations in endothelial barrier function in disturbed flows in vitro. , 2000, American journal of physiology. Heart and circulatory physiology.

[11]  R. Ian Freshney,et al.  Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications , 2010 .

[12]  D E H Tee,et al.  Book Review: Culture of Animal Cells: A Manual of Basic Technique , 1984 .

[13]  M. Chagnon,et al.  Uridine uptake inhibition as a cytotoxicity test for a human hepatoma cell line (HepG2 cells): comparison with the neutral red assay. , 2001, Toxicology.

[14]  M. Grant,et al.  Immortalized hepatocytes as in vitro model systems for toxicity testing: the comparative toxicity of menadione in immortalized cells, primary cultures of hepatocytes and HTC hepatoma cells. , 1996, Toxicology in vitro : an international journal published in association with BIBRA.

[15]  Andreas Janshoff,et al.  Cell adhesion monitoring using a quartz crystal microbalance: comparative analysis of different mammalian cell lines , 1998, European Biophysics Journal.

[16]  Joachim Wegener,et al.  Real-time impedance assay to follow the invasive activities of metastatic cells in culture. , 2002, BioTechniques.

[17]  P. Cochard,et al.  Dual staining assessment of Schwann cell viability within whole peripheral nerves using calcein-AM and ethidium homodimer , 1997, Journal of Neuroscience Methods.

[18]  B Wolf,et al.  Monitoring of cellular signalling and metabolism with modular sensor-technique: the PhysioControl-Microsystem (PCM). , 1998, Biosensors & bioelectronics.

[19]  R. E. White,et al.  High-throughput screening in drug metabolism and pharmacokinetic support of drug discovery. , 2000, Annual review of pharmacology and toxicology.

[20]  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.

[21]  C.R. Keese,et al.  A biosensor that monitors cell morphology with electrical fields , 1994, IEEE Engineering in Medicine and Biology Magazine.

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

[23]  M. D. Rooij,et al.  Electrochemical Methods: Fundamentals and Applications , 2003 .

[24]  G. R. Facer,et al.  Capacitance cytometry: measuring biological cells one by one. , 2000, Proceedings of the National Academy of Sciences of the United States of America.