Real-time electrical impedance-based measurement to distinguish oral cancer cells and non-cancer oral epithelial cells

In this study, electrical impedance-based measurements were used to distinguish oral cancer cells and non-cancer oral epithelial cells based on their cellular activities on the microelectrodes in a real-time and label-free manner. CAL 27 and Het-1A cell lines were used as the models of oral cancer cells and non-cancer oral epithelial cells, respectively. Various cellular activities, including cell adhesion, spreading, and proliferation were monitored. We found that both the kinetics of cell spreading and the static impedance-based cell index were feasible to distinguish the two cell types. At each given cell number, CAL 27 cell spreading produced a smaller cell index change rate that was 60–70% of those of Het-1A cells. When cells were fully spread, CAL 27 cells generated a cell index more than four times greater than that of Het-1A cells. Since cell spreading and attachment occurs in the first few hours when they were cultured on the microelectrodes, this impedance-based method could be a rapid label-free and non-invasive approach to distinguish oral cancer cells from non-cancer oral epithelial cells. Cell viability analysis was performed along with the impedance-based analysis. Confocal microscopic imaging analysis showed the difference in cell morphology and the thickness of cell monolayers between the two cell types.

[1]  H. Schwan.  Electrical properties of tissue and cell suspensions. , 1957, Advances in biological and medical physics.

[2]  A. Schwinde,et al.  On-line control of cellular adhesion with impedance measurements using interdigitated electrode structures , 1998, Medical and Biological Engineering and Computing.

[3]  Richard D. Rabbitt,et al.  Electric impedance spectroscopy using microchannels with integrated metal electrodes , 1999 .

[4]  A. B. Frazier,et al.  Microsystems for isolation and electrophysiological analysis of breast cancer cells from blood. , 2006, Biosensors & bioelectronics.

[5]  Benjamin Geiger,et al.  Focal Contacts as Mechanosensors Externally Applied Local Mechanical Force Induces Growth of Focal Contacts by an Mdia1-Dependent and Rock-Independent Mechanism , 2001 .

[6]  Michael P Hughes,et al.  Early detection of oral cancer - Is dielectrophoresis the answer? , 2007, Oral oncology.

[7]  Dielectric measurement of a single sub-millimeter size microcapsule , 1994 .

[8]  Boris Rubinsky,et al.  A novel microfluidic cell-chip for single cell analysis and manipulation , 2003, The Sixteenth Annual International Conference on Micro Electro Mechanical Systems, 2003. MEMS-03 Kyoto. IEEE.

[9]  Giljun Park,et al.  Electrical impedance measurements predict cellular transformation , 2009, Cell biology international.

[10]  L. Addadi,et al.  Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates , 2001, Nature Cell Biology.

[11]  D E Ingber,et al.  Vinculin promotes cell spreading by mechanically coupling integrins to the cytoskeleton. , 1997, Experimental cell research.

[12]  M. Weinberg,et al.  Assessing oral malignancies. , 2002, American family physician.

[13]  R. Cote,et al.  Detection and clinical importance of micrometastatic disease. , 1999, Journal of the National Cancer Institute.

[14]  T. Day,et al.  Oral Cancer and Precancerous Lesions , 2002, CA: a cancer journal for clinicians.

[15]  Michael Fischer,et al.  Microcavity array (MCA)-based biosensor chip for functional drug screening of 3D tissue models. , 2008, Biosensors & bioelectronics.

[16]  Ivar Giaever,et al.  TOXIC ? CELLS CAN TELL , 1992 .

[17]  L. Zhao,et al.  Transcriptional expression profiles of oral squamous cell carcinomas , 2002, Cancer.

[18]  C. Dawes,et al.  Estimates, from salivary analyses, of the turnover time of the oral mucosal epithelium in humans and the number of bacteria in an edentulous mouth. , 2003, Archives of oral biology.

[19]  Qingjun Liu,et al.  Impedance studies of bio-behavior and chemosensitivity of cancer cells by micro-electrode arrays. , 2009, Biosensors & bioelectronics.

[20]  William E. Ortyn,et al.  Cellular image analysis and imaging by flow cytometry. , 2007, Clinics in laboratory medicine.

[21]  G. Whitesides,et al.  Cell shape provides global control of focal adhesion assembly. , 2003, Biochemical and biophysical research communications.

[22]  Martin A. Schwartz,et al.  Networks and crosstalk: integrin signalling spreads , 2002, Nature Cell Biology.

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

[24]  Yanping Wang,et al.  Real-time monitoring approach: assessment of effects of antibodies on the adhesion of NCI-H460 cancer cells to the extracellular matrix. , 2008, Biosensors & bioelectronics.

[25]  S. Gawad,et al.  Impedance Spectroscopy Cell Analysis in Microchannels , 2001 .

[26]  N. Balaban,et al.  Calculation of forces at focal adhesions from elastic substrate data: the effect of localized force and the need for regularization. , 2002, Biophysical journal.

[27]  A. B. Frazier,et al.  A micro system using dielectrophoresis and electrical impedance spectroscopy for cell manipulation and analysis , 2003, TRANSDUCERS '03. 12th International Conference on Solid-State Sensors, Actuators and Microsystems. Digest of Technical Papers (Cat. No.03TH8664).

[28]  Donald E Ingber,et al.  Micropatterning tractional forces in living cells. , 2002, Cell motility and the cytoskeleton.

[29]  Liju Yang,et al.  Real-time electrical impedance detection of cellular activities of oral cancer cells. , 2010, Biosensors & bioelectronics.

[30]  S. Takashima,et al.  Frequency domain studies of impedance characteristics of biological cells using micropipet technique. I. Erythrocyte. , 1988, Biophysical journal.

[31]  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.

[32]  R. McGuinness.  Impedance-based cellular assay technologies: recent advances, future promise. , 2007, Current opinion in pharmacology.

[33]  Jian Xu,et al.  Response characteristics of single-cell impedance sensors employed with surface-modified microelectrodes. , 2010, Biosensors & bioelectronics.

[34]  Artur Ferreira,et al.  Oral squamous cell carcinoma: review of prognostic and predictive factors. , 2006, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[35]  R. Orlando,et al.  Regulatory volume decrease in human esophageal epithelial cells. , 2002, American journal of physiology. Gastrointestinal and liver physiology.

[36]  Christopher S. Chen,et al.  Cells lying on a bed of microneedles: An approach to isolate mechanical force , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[37]  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.

[38]  Pontus Linderholm,et al.  Bipolar resistivity profiling of 3D tissue culture. , 2007, Biosensors & bioelectronics.

[39]  L. Mcdonald,et al.  The pathology of oral cancer , 2003, Pathology.

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

[41]  M. Bissell,et al.  Extracellular matrix signaling: integration of form and function in normal and malignant cells. , 1998, Current opinion in cell biology.

[42]  C. Harris,et al.  Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. , 1991, Cancer research.

[43]  S. Hanks,et al.  Induced Focal Adhesion Kinase (FAK) Expression in FAK-Null Cells Enhances Cell Spreading and Migration Requiring Both Auto- and Activation Loop Phosphorylation Sites and Inhibits Adhesion-Dependent Tyrosine Phosphorylation of Pyk2 , 1999, Molecular and Cellular Biology.

[44]  Ivar Giaever,et al.  A morphological biosensor for mammalian cells , 1993, Nature.

[45]  G. Boeck,et al.  Current status of flow cytometry in cell and molecular biology. , 2001, International review of cytology.

[46]  Jian Xu,et al.  Influence of cell adhesion and spreading on impedance characteristics of cell-based sensors. , 2008, Biosensors & bioelectronics.

[47]  Zongyuan Chen,et al.  Lab‐on‐a‐Chip Technologies for Oral‐Based Cancer Screening and Diagnostics , 2007, Annals of the New York Academy of Sciences.

[48]  I. Giaever,et al.  Micromotion of mammalian cells measured electrically. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Voldman,et al.  Dielectrophoretic registration of living cells to a microelectrode array. , 2004, Biosensors & bioelectronics.