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

Ilhwan, Park; Hong, Yeonhee; Jun, Young Hoo; Lee, Ga Yeon; Jun, Hee-Sook; Pyun, Jae Chul; Choi, Jeong Woo; Cho, Sungbo

doi:10.3390/s16122068

Cited by 13 publications

(12 citation statements)

References 49 publications

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“…To overcome hurdles in studying myogenesis, other tools have been designed to assess myoblast differentiation in real-time based on live cell monitoring. These tools include assessment of physical properties and function such as contractility through displacement analysis [ 36 ], skeletal muscle thickness [ 45 , 46 ] and myoblast differentiation [ 46 ] through electrical impedance. Additionally, multiple imaging modalities have been applied to assess muscle biology, including bioluminescence imaging for luciferase tagged cell transplantation [ 47 ], second harmonic generation (SHG) imaging to quantify muscle striation patterns [ 48 , 49 ], MRI to assess oxidative phosphorylation in muscle [ 50 ], and multi-photon imaging for the assessment of myoblast oxidation level [ 51 ].…”

Section: Discussionmentioning

confidence: 99%

A real-time monitoring platform of myogenesis regulators using double fluorescent labeling

Niu¹,

Zhou²,

Prim³

et al. 2018

PLoS ONE

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Real-time, quantitative measurement of muscle progenitor cell (myoblast) differentiation is an important tool for skeletal muscle research and identification of drugs that support skeletal muscle regeneration. While most quantitative tools rely on sacrificial approach, we developed a double fluorescent tagging approach, which allows for dynamic monitoring of myoblast differentiation through assessment of fusion index and nuclei count. Fluorescent tagging of both the cell cytoplasm and nucleus enables monitoring of cell fusion and the formation of new myotube fibers, similar to immunostaining results. This labeling approach allowed monitoring the effects of Myf5 overexpression, TNFα, and Wnt agonist on myoblast differentiation. It also enabled testing the effects of surface coating on the fusion levels of scaffold-seeded myoblasts. The double fluorescent labeling of myoblasts is a promising technique to visualize even minor changes in myogenesis of myoblasts in order to support applications such as tissue engineering and drug screening.

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Section: Discussionmentioning

confidence: 99%

A real-time monitoring platform of myogenesis regulators using double fluorescent labeling

Niu¹,

Zhou²,

Prim³

et al. 2018

PLoS ONE

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“…The ITO electrode, which was established in our previous studies [22,26], was prepared as shown in Figure 1. The brief fabrication process of the ITO electrode-based cultureware is as follows.…”

Section: Fabrication Of the Ito Electrode-based Culturewarementioning

confidence: 99%

“…For the impedance monitoring of cells, the established experimental setup [22,26] was utilized, which consisted of the digital lock-in-amplifier (SR830; Stanford Research Systems, Sunnyvale, CA, USA), multiplexer, and LabVIEW program. The amplitude of the current flowing through the cells on the electrode was restricted to below 1 µA to minimize electrical effects on the cells.…”

Section: Impedance Measurement Of Cells On the Rgo-aunp/ito Electrodementioning

confidence: 99%

“…The transparent indium tin oxide (ITO) electrode made it possible to characterize the impedance of cells with better optical transmittance when compared to typical gold or platinum electrodes [20,21]. The surface of the ITO electrode could be modified via protein, peptide, or organic polymer coatings to achieve the biocompatibility [22][23][24][25] required for the impedance monitoring of cells. However, organic materials are easily affected by environmental factors including temperature, humidity, pH, and solvent ions, and can be denatured such that substrate conditions cannot guarantee cell cultivation.…”

Section: Introductionmentioning

confidence: 99%

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Electrochemical Impedance Characterization of Cell Growth on Reduced Graphene Oxide–Gold Nanoparticles Electrodeposited on Indium Tin Oxide Electrodes

et al. 2019

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The improved binding ability of graphene–nanoparticle composites to proteins or molecules can be utilized to develop new cell-based assays. In this study, we fabricated reduced graphene oxide–gold nanoparticles (rGO-AuNP) electrodeposited onto a transparent indium tin oxide (ITO) electrode and investigated the feasibility of the electrochemical impedance monitoring of cell growth. The electrodeposition of rGO–AuNP on the ITO was optically and electrochemically characterized in comparison to bare, rGO-, and AuNP-deposited electrodes. The cell growth on the rGO–AuNP/ITO electrode was analyzed via electrochemical impedance measurement together with the microscopic observation of HEK293 cells transfected with a green fluorescent protein expression vector. The results showed that rGO–AuNP was biocompatible and induced an increase in cell adherence to the electrode when compared to the bare, AuNP-, or rGO-deposited ITO electrode. At 54 h cultivation, the average and standard deviation of the saturated normalized impedance magnitude of the rGO–AuNP/ITO electrode was 3.44 ± 0.16, while the value of the bare, AuNP-, and rGO-deposited ITO electrode was 2.48 ± 0.15, 2.61 ± 0.18, and 3.01 ± 0.25, respectively. The higher saturated value of the cell impedance indicates that the impedimetric cell-based assay has a broader measurement range. Thus, the rGO–AuNP/ITO electrode can be utilized for label-free and real-time impedimetric cell-based assays with wider dynamic range.

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“…Muscle cells respond to electric fields and induce the contraction of muscle tissue. Thus, the local electric field generated by ZnO NSs would potentially improve the regeneration and rehabilitation of muscle tissue, with several future applications in chronic illnesses such as muscle atrophy, wasting and aging [ 19 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

Biocompatibility and Electrical Stimulation of Skeletal and Smooth Muscle Cells Cultured on Piezoelectric Nanogenerators

Blanquer

Careta

Anido-Varela

et al. 2021

IJMS

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Nanogenerators are interesting for biomedical applications, with a great potential for electrical stimulation of excitable cells. Piezoelectric ZnO nanosheets present unique properties for tissue engineering. In this study, nanogenerator arrays based on ZnO nanosheets are fabricated on transparent coverslips to analyse the biocompatibility and the electromechanical interaction with two types of muscle cells, smooth and skeletal. Both cell types adhere, proliferate and differentiate on the ZnO nanogenerators. Interestingly, the amount of Zn ions released over time from the nanogenerators does not interfere with cell viability and does not trigger the associated inflammatory response, which is not triggered by the nanogenerators themselves either. The local electric field generated by the electromechanical nanogenerator–cell interaction stimulates smooth muscle cells by increasing cytosolic calcium ions, whereas no stimulation effect is observed on skeletal muscle cells. The random orientation of the ZnO nanogenerators, avoiding an overall action potential aligned along the muscle fibre, is hypothesised to be the cause of the cell-type dependent response. This demonstrates the need of optimizing the nanogenerator morphology, orientation and distribution according to the potential biomedical use. Thus, this study demonstrates the cell-scale stimulation triggered by biocompatible piezoelectric nanogenerators without using an external source on smooth muscle cells, although it remarks the cell type-dependent response.

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

Cited by 13 publications

References 49 publications

A real-time monitoring platform of myogenesis regulators using double fluorescent labeling

A real-time monitoring platform of myogenesis regulators using double fluorescent labeling

Electrochemical Impedance Characterization of Cell Growth on Reduced Graphene Oxide–Gold Nanoparticles Electrodeposited on Indium Tin Oxide Electrodes

Biocompatibility and Electrical Stimulation of Skeletal and Smooth Muscle Cells Cultured on Piezoelectric Nanogenerators

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