In vitro hyperthermia studied in a continuous manner using electric impedance sensing

In this study, a new platform based on electric cell-substrate impedance sensing (ECIS) was constructed for the dynamic monitoring of changes in cells during and after hyperthermia treatments. ECIS profiling was compared with traditional methods for monitoring the status of A549 cells under three typical treatment conditions, i.e., 30 min of hyperthermia at 41, 43, and 45 °C. The impedance value rapidly changed, and severe morphological changes were observed during and after the hyperthermia. The impedance curves revealed that different hyperthermia conditions differentially affected the cells: the 41 °C treatment caused a minor decrease in impedance that almost completely recovered in 1–2 h; the 43 °C treatment led to a greater decrease in impedance, which also recovered over several hours before slowly decreasing again, possibly indicating apoptosis; the 45 °C treatment resulted in the greatest decrease in impedance, which never recovered, possibly indicating rapid necrosis. Further, these three hyperthermia treatment regimens were applied to four additional cell lines. By comparing the impedance curves of different cell lines, we found that cancer cells (HepG2) may be more sensitive to hyperthermia than normal cells (LO2). Moreover, different cancer cell lines (HeLa, MCF-7, A549, and HepG2) exhibited different thermal sensitivities. These results fit previous theories on hyperthermia, demonstrating that the platform established in this study is a useful analytical tool for the in vitro research of thermal therapy, and the dynamic data generated will enable us to examine phenomena and theories.

[1]  R. Issels Hyperthermia adds to chemotherapy. , 2008, European journal of cancer.

[2]  Kevin Barraclough,et al.  I and i , 2001, BMJ : British Medical Journal.

[3]  J. L. Roti,et al.  Cellular responses to hyperthermia (40-46°C) : Cell killing and molecular events , 2008 .

[4]  R. Dirksen,et al.  Temperature and RyR1 regulate the activation rate of store-operated Ca²+ entry current in myotubes. , 2012, Biophysical journal.

[5]  Reyes Sierra-Alvarez,et al.  Application and validation of an impedance-based real time cell analyzer to measure the toxicity of nanoparticles impacting human bronchial epithelial cells. , 2012, Environmental science & technology.

[6]  Lei Wang,et al.  Use of cellular electrical impedance sensing to assess in vitro cytotoxicity of anticancer drugs in a human kidney cell nephrotoxicity model. , 2012, The Analyst.

[7]  J. Bull,et al.  Apoptosis in tumors and normal tissues induced by whole body hyperthermia in rats. , 1995, Cancer research.

[8]  G. Ávila,et al.  Distinct effects on Ca2+ handling caused by malignant hyperthermia and central core disease mutations in RyR1. , 2004, Biophysical journal.

[9]  Jing Zhu,et al.  Real-time monitoring of extracellular matrix-mediated PC12 cell attachment and proliferation using an electronic biosensing device , 2011, Biotechnology Letters.

[10]  Tien Anh Nguyen,et al.  Microfluidic chip with integrated electrical cell-impedance sensing for monitoring single cancer cell migration in three-dimensional matrixes. , 2013, Analytical chemistry.

[11]  J. Pirro,et al.  The response of human and rodent cells to hyperthermia. , 1991, International journal of radiation oncology, biology, physics.

[12]  Sanghyo Kim,et al.  Impedance-based cell culture platform to assess light-induced stress changes with antagonist drugs using retinal cells. , 2013, Analytical chemistry.

[13]  Aaas News,et al.  Book Reviews , 1893, Buffalo Medical and Surgical Journal.

[14]  L. Bourget,et al.  Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis , 1997, Molecular and cellular biology.

[15]  Jing Cheng,et al.  Bioelectrical Impedance Assay to Monitor Changes in Aspirin‐Treated Human Colon Cancer HT‐29 Cell Shape during Apoptosis , 2007 .

[16]  Zeljko Vujaskovic,et al.  Randomized trial of hyperthermia and radiation for superficial tumors. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[17]  T. Otsuka,et al.  Hyperthermia induces apoptosis in malignant fibrous histiocytoma cells in vitro , 1996, International journal of cancer.

[18]  Janusz Skowronek,et al.  Hyperthermia – description of a method and a review of clinical applications , 2007 .

[19]  David Schneider,et al.  Cytotoxicity of metal and semiconductor nanoparticles indicated by cellular micromotility. , 2009, ACS nano.

[20]  Mark W. Dewhirst,et al.  Prospective thermal dosimetry: The key to hyperthermia's future , 2006, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[21]  Jintian Tang,et al.  Effect of hyperthermia on the apoptosis and proliferation of CaSki cells. , 2010, Molecular medicine reports.

[22]  W. Dewey,et al.  Arrhenius relationships from the molecule and cell to the clinic , 2009, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[23]  Lei Wang,et al.  Real-time, label-free monitoring of the cell cycle with a cellular impedance sensing chip. , 2010, Biosensors & bioelectronics.

[24]  H. C. Mastwijk,et al.  Electroporation of cells in microfluidic devices: a review , 2006, Analytical and bioanalytical chemistry.

[25]  R. Kopelman,et al.  Photothermal therapy of cancer cells mediated by blue hydrogel nanoparticles. , 2012, Nanomedicine.

[26]  Jing Zhu,et al.  An automatic and quantitative on-chip cell migration assay using self-assembled monolayers combined with real-time cellular impedance sensing. , 2008, Lab on a chip.

[27]  J C Bischof,et al.  In vitro thermal therapy of AT-1 Dunning prostate tumours , 2004, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[28]  J. Kerr,et al.  Cell death induced in a murine mastocytoma by 42-47 degrees C heating in vitro: evidence that the form of death changes from apoptosis to necrosis above a critical heat load. , 1990, International journal of radiation biology.

[29]  Sungbo Cho,et al.  Hydrogel-based diffusion chip with Electric Cell-substrate Impedance Sensing (ECIS) integration for cell viability assay and drug toxicity screening. , 2013, Biosensors & bioelectronics.

[30]  P. Wust,et al.  The cellular and molecular basis of hyperthermia. , 2002, Critical reviews in oncology/hematology.

[31]  C. Voermans,et al.  BIGH3 modulates adhesion and migration of hematopoietic stem and progenitor cells , 2013, Cell adhesion & migration.

[32]  W. Marsden I and J , 2012 .

[33]  Ali Khademhosseini,et al.  Study of long-term viability of endothelial cells for lab-on-a-chip devices , 2013 .

[34]  D. S. Coffey,et al.  Hyperthermic biology and cancer therapies: a hypothesis for the "Lance Armstrong effect". , 2006, JAMA.

[35]  H. A. Schwettman,et al.  Cellular tolerance to pulsed hyperthermia. , 2006, Physical review. E, Statistical, nonlinear, and soft matter physics.

[36]  C. Lim,et al.  Cell cycle dependent apoptosis and cell cycle blocks induced by hyperthermia in HL-60 cells , 2006, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[37]  P. J. Hoopes,et al.  Basic principles of thermal dosimetry and thermal thresholds for tissue damage from hyperthermia , 2003, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[38]  Pierre O. Bagnaninchi,et al.  Real-time label-free monitoring of adipose-derived stem cell differentiation with electric cell-substrate impedance sensing , 2011, Proceedings of the National Academy of Sciences.

[39]  Joachim Wegener,et al.  Impedance analysis of adherent cells after in situ electroporation: non-invasive monitoring during intracellular manipulations. , 2011, Biosensors & bioelectronics.

[40]  Jing Cheng,et al.  Real-Time, Label-Free Monitoring of 3T3-L1 Preadipocyte Differentiation by a Bioelectrical Impedance Assay , 2012 .

[41]  C. Lo,et al.  Impedance analysis of MDCK cells measured by electric cell-substrate impedance sensing. , 1995, Biophysical journal.

[42]  Lei Wang,et al.  A microfluidic device with passive air-bubble valves for real-time measurement of dose-dependent drug cytotoxicity through impedance sensing. , 2012, Biosensors & bioelectronics.

[43]  R. Mahajan,et al.  Hyperthermia induces endoplasmic reticulum-mediated apoptosis in melanoma and non-melanoma skin cancer cells. , 2008, The Journal of investigative dermatology.

[44]  Lei Wang,et al.  Analysis of the sensitivity and frequency characteristics of coplanar electrical cell-substrate impedance sensors. , 2008, Biosensors & bioelectronics.

[45]  Ching-Te Huang,et al.  Supraphysiological Thermal Injury in Different Human Bladder Carcinoma Cell Lines , 2009, Annals of Biomedical Engineering.