Electrical Impedance Monitoring of C2C12 Myoblast Differentiation on an Indium Tin Oxide Electrode

Electrical cell-substrate impedance sensing is increasingly being used for label-free and real-time monitoring of changes in cell morphology and number during cell growth, drug screening, and differentiation. In this study, we evaluated the feasibility of using ECIS to monitor C2C12 myoblast differentiation using a fabricated indium tin oxide (ITO) electrode-based chip. C2C12 myoblast differentiation on the ITO electrode was validated based on decreases in the mRNA level of MyoD and increases in the mRNA levels of myogenin and myosin heavy chain (MHC). Additionally, MHC expression and morphological changes in myoblasts differentiated on the ITO electrode were comparable to those in cells in the control culture dish. From the monitoring the integration of the resistance change at 21.5 kHz, the cell differentiation was label-free and real-time detectable in 30 h of differentiation (p < 0.05).

[1]  Joachim Wegener,et al.  Electrical wound-healing assay for cells in vitro. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Jeong-Woo Choi,et al.  In situ electrochemical detection of embryonic stem cell differentiation. , 2013, Journal of biotechnology.

[3]  P. Jakeman,et al.  Optimization of an in vitro bioassay to monitor growth and formation of myotubes in real time , 2016, Bioscience reports.

[4]  Sungbo Cho,et al.  Detection of the osteogenic differentiation of mesenchymal stem cells in 2D and 3D cultures by electrochemical impedance spectroscopy. , 2010, Journal of biotechnology.

[5]  S. Kuang,et al.  Myostatin facilitates slow and inhibits fast myosin heavy chain expression during myogenic differentiation. , 2012, Biochemical and biophysical research communications.

[6]  Sang-Jin Lee,et al.  Bakuchiol augments MyoD activation leading to enhanced myoblast differentiation. , 2016, Chemico-biological interactions.

[7]  H. Tsutsui,et al.  Skeletal Muscle Abnormalities in Heart Failure. , 2015, International heart journal.

[8]  Xin Zhang,et al.  Real-time monitoring primary cardiomyocyte adhesion based on electrochemical impedance spectroscopy and electrical cell-substrate impedance sensing. , 2008, Analytical chemistry.

[9]  A. Edkins,et al.  Real-time monitoring of 3T3-L1 preadipocyte differentiation using a commercially available electric cell-substrate impedance sensor system. , 2014, Biochemical and biophysical research communications.

[10]  D. Scaini,et al.  In vitro myogenesis induced by human recombinant elastin-like proteins. , 2015, Biomaterials.

[11]  S. Rakhilin,et al.  Electrical Impedance as a Novel Biomarker of Myotube Atrophy and Hypertrophy , 2011, Journal of biomolecular screening.

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

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

[14]  Xin Zhang,et al.  Impedance-based monitoring of ongoing cardiomyocyte death induced by tumor necrosis factor-alpha. , 2009, Biophysical journal.

[15]  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.

[16]  Jeong-Woo Choi,et al.  3D graphene oxide-encapsulated gold nanoparticles to detect neural stem cell differentiation. , 2013, Biomaterials.

[17]  Sungbo Cho Electrical impedance simulation and characterization of cell growth using the Fricke model. , 2012, Journal of nanoscience and nanotechnology.

[18]  J. Ryall The role of sirtuins in the regulation of metabolic homeostasis in skeletal muscle , 2012, Current opinion in clinical nutrition and metabolic care.

[19]  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.

[20]  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.

[21]  J. Min,et al.  Indium tin oxide based chip for optical and electrochemical characterization of protein–cell interaction , 2015 .

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

[23]  S. Lindstedt,et al.  Skeletal muscle tissue in movement and health: positives and negatives , 2016, Journal of Experimental Biology.

[24]  Sungbo Cho,et al.  Electrical impedance characterization of cell growth on interdigitated microelectrode array. , 2014, Journal of Nanoscience and Nanotechnology.

[25]  T. Hawke,et al.  Diabetic myopathy: impact of diabetes mellitus on skeletal muscle progenitor cells , 2013, Front. Physiol..

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

[27]  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.

[28]  S. Brochsztain,et al.  Characterization of a Perylenediimide Self-Assembled Monolayer on Indium Tin Oxide Electrodes Using Electrochemical Impedance Spectroscopy , 2014 .

[29]  A. Russell,et al.  Skeletal muscle mitochondria: a major player in exercise, health and disease. , 2014, Biochimica et biophysica acta.

[30]  Alberto Olmo,et al.  Monitoring living cell assays with bio-impedance sensors , 2013 .

[31]  B. Staels,et al.  Skeletal muscle functions around the clock , 2015, Diabetes, obesity & metabolism.

[32]  Szi-Wen Chen,et al.  A computational modeling and analysis in cell biological dynamics using electric cell-substrate impedance sensing (ECIS). , 2012, Biosensors & bioelectronics.

[33]  Yonghyun Choi,et al.  Electrochemical Characterization of Poly-L-Lysine Coating on Indium Tin Oxide Electrode for Enhancing Cell Adhesion. , 2015, Journal of nanoscience and nanotechnology.

[34]  Sungbo Cho,et al.  Electrical characterization of human mesenchymal stem cell growth on microelectrode , 2008 .

[35]  Szi-Wen Chen,et al.  A quantitative cell modeling and wound-healing analysis based on the Electric Cell-substrate Impedance Sensing (ECIS) method , 2016, Comput. Biol. Medicine.

[36]  Sungbo Cho,et al.  Chip-based time-continuous monitoring of toxic effects on stem cell differentiation. , 2009, Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft.

[37]  Letao Yang,et al.  Controlling differentiation of adipose-derived stem cells using combinatorial graphene hybrid-pattern arrays. , 2015, ACS nano.

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

[39]  Lan T M Dao,et al.  Effect of cell senescence on the impedance measurement of adipose tissue-derived stem cells. , 2013, Enzyme and microbial technology.

[40]  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.

[41]  Gun Young Jung,et al.  Large‐Scale Nanoelectrode Arrays to Monitor the Dopaminergic Differentiation of Human Neural Stem Cells , 2015, Advanced materials.

[42]  Sungbo Cho,et al.  Real-Time Monitoring of Neural Differentiation of Human Mesenchymal Stem Cells by Electric Cell-Substrate Impedance Sensing , 2011, Journal of biomedicine & biotechnology.

[43]  Mi-Ok Lee,et al.  In situ label-free quantification of human pluripotent stem cells with electrochemical potential. , 2016, Biomaterials.

[44]  M. Gajewska,et al.  FOXO1 and GSK-3β Are Main Targets of Insulin-Mediated Myogenesis in C2C12 Muscle Cells , 2016, PloS one.

[45]  V. Aas,et al.  Electrical Stimulation Improves Insulin Responses in a Human Skeletal Muscle Cell Model of Hyperglycemia , 2002, Annals of the New York Academy of Sciences.

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