Study of Cells Attachment Using Impedance Spectroscopy Technique

2011 ◽  
Author(s):  
Anis N. Nordin ◽  
S. M. Arifuzzaman ◽  
Maizirwan Mel ◽  
David Spray ◽  
Ioana Voiculescu
Keyword(s):  
Cell Culture ◽  
Endothelial Cells ◽  
Impedance Spectroscopy ◽  
Culture Medium ◽  
Equivalent Circuit Model ◽  
Electrical Impedance Spectroscopy ◽  
Fibroblast Cells ◽  
Impedance Sensing ◽  
Impedance Measurements ◽  
Mammalian Cell Lines

In this paper we report on the electrical impedance spectroscopy characterization of the attachment of two mammalian cell lines: endothelial cells (RFPEC) and fibroblast cells (V79). For electrical cell–substrate impedance sensing (ECIS) of the endothelial cells a commercially available eight-well cell culture impedance array (ECIS-8W1E) was used. The impedance measurements were recorded with cell culture medium (without cells) and with endothelial cells or fibroblast cells in the culture medium over a frequency range from 100 Hz to 100 kHz. The impedance measurements were compared to the equivalent circuit model. The impedance measurements were also simulated using COMSOL Multiphysics™, a commercially available modeling package.

scholarly journals Effects of Lead Exposure on Blood Electrical Impedance Spectroscopy of Mice

2021 ◽  
Author(s):  
Binying Yang ◽  
Jia Xu ◽  
Shao Hu ◽  
Boning You ◽  
Qing Ma
Keyword(s):  
Electrical Properties ◽  
Impedance Spectroscopy ◽  
Lead Exposure ◽  
Electrical Impedance ◽  
Equivalent Circuit Model ◽  
Impedance Spectrum ◽  
Nyquist Plot ◽  
Electrical Impedance Spectroscopy ◽  
Impedance Method ◽  
Range Data

Abstract Background: Lead is a nonessential heavy metal, which can inhibit heme synthesis and has significant cytotoxic effects. Nevertheless, its effect on the electrical properties of red blood cells (RBCs) remains unclear. Consequently, this study aimed to investigate the electrical properties and the electrophysiological mechanism of lead exposure in mouse blood using Electrical Impedance Spectroscopy (EIS). Methods: AC impedance method was used to measure the electrical impedance of healthy and lead exposure blood of mice in 0.01-100 MHz frequency range. Data characteristic of the impedance spectrum, Bodes plot, Nyquist plot and Nichols plot, and three elements equivalent circuit model were used to explicitly analyze the differences in amplitude-frequency, phase-frequency, and the frequency characteristic of blood in electrical impedance properties. Results: Compared with the healthy blood in control mice, the changes in blood exposed to lead was as follows: (I) the hematocrit decreased; (II) the amplitude-frequency and phase-frequency characteristics of electrical impedance decreased; (III) the characteristic frequencies ( f 0 ) were significantly increased; (IV) the electrical impedance of plasma, erythrocyte membrane, and hemoglobin decreased, while the conductivity increased. Conclusion: Therefore, EIS can be used as an effective method to monitor blood and RBCs abnormalities caused by lead-exposure.

Endothelial cell cystine uptake and glutathione increase with N,N-bis(2-chloroethyl)-N-nitrosourea exposure

1992 ◽  
Vol 262 (3) ◽  
pp. L301-L304 ◽  
Author(s):  
S. M. Deneke ◽  
R. A. Lawrence ◽  
S. G. Jenkinson
Keyword(s):  
Cell Culture ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Culture Medium ◽  
Cultured Cells ◽  
Potent Inhibitor ◽  
Cell Types ◽  
Cell Culture Medium ◽  
Intracellular Accumulation ◽  
Pulmonary Endothelial Cells

Glutathione (gamma-glutamylcysteinylglycine, GSH) is an important cellular antioxidant. In typical cultured cell preparations GSH synthesis is limited by the availability of intracellular cysteine. Because extracellular cystine is the chief source of intracellular cysteine in cultured cells, increasing cystine transport can result in increased intracellular GSH. Depletion of GSH or exposure to oxidants has been shown to stimulate cystine transport in bovine pulmonary endothelial cells and other cell types. BCNU [N,N-bis(2-chloroethyl)-N-nitrosourea] is a potent inhibitor of glutathione reductase (GSSG-Red). We examined the effects of BCNU on cystine uptake by bovine pulmonary artery endothelial cells (BPAEC). We hypothesized that blocking GSSG-Red could result in increased cellular uptake of cystine to replenish decreases in GSH caused by oxidation. Levels of BCNU between 0.005 and 0.05 mM added to the cell culture medium inhibited GSSG-Red at 2, 4, and 24 h after addition. BCNU treatment resulted in concentration-dependent increases in both cystine uptake and GSH levels after 24 h of exposure. The increases in uptake were specific for cystine and glutamate and were sodium independent, suggesting induction of a xc(-)-like transport system. No intracellular accumulation of GSSG was measured nor was any significant depletion of GSH noted at any time of BCNU exposure.

scholarly journals Addition of Exogenous Protease Facilitates Reovirus Infection in Many Restrictive Cells

2002 ◽  
Vol 76 (15) ◽  
pp. 7430-7443 ◽  
Author(s):  
Joseph W. Golden ◽  
Jessica Linke ◽  
Stephen Schmechel ◽  
Kara Thoemke ◽  
Leslie A. Schiff
Keyword(s):  
Cell Culture ◽  
Life Cycle ◽  
Host Range ◽  
Cell Lines ◽  
Culture Medium ◽  
Cathepsin L ◽  
Cell Culture Medium ◽  
Reovirus Infection ◽  
Lysosomal Protease ◽  
Mammalian Cell Lines

ABSTRACT Virion uncoating is a critical step in the life cycle of mammalian orthoreoviruses. In cell culture, and probably in extraintestinal tissues in vivo, reovirus virions undergo partial proteolysis within endosomal or/or lysosomal compartments. This process converts the virion into a form referred to as an intermediate subvirion particle (ISVP). In natural enteric reovirus infections, proteolytic uncoating takes place extracellularly within the intestinal lumen. The resultant proteolyzed particles, unlike intact virions, have the capacity to penetrate cell membranes and thereby gain access to cytoplasmic components required for viral gene expression. We hypothesized that the capacity of reovirus outer capsid proteins to be proteolyzed is a determinant of cellular host range. To investigate this hypothesis, we asked if the addition of protease to cell culture medium would expand the range of cultured mammalian cell lines that can be productively infected by reoviruses. We identified many transformed and nontransformed cell lines, as well as primary cells, that restrict viral infection. In several of these restrictive cells, virion uncoating is inefficient or blocked. Addition of proteases to the cell culture medium generates ISVP-like particles and promotes viral growth in nearly all cell lines tested. Interestingly, we found that some cell lines that restrict reovirus uncoating still express mature cathepsin L, a lysosomal protease required for virion disassembly in murine L929 cells. This finding suggests that factors in addition to cathepsin L are required for efficient intracellular proteolysis of reovirus virions. Our results demonstrate that virion uncoating is a critical determinant of reovirus cellular host range and that many cells which otherwise support productive reovirus infection cannot efficiently mediate this essential early step in the virus life cycle.

scholarly journals Proliferative Effects of Nitrogen Nanobubbles on Fibroblasts

2021 ◽  
Author(s):  
Hansol Heo ◽  
Junseon Park ◽  
Jeong Il Lee ◽  
Jungho Kim ◽  
Joong Yull Park ◽  
...  
Keyword(s):  
Cell Culture ◽  
Flow Cytometry ◽  
Cell Proliferation ◽  
Image Analysis ◽  
Culture Medium ◽  
Fibroblast Cells ◽  
High Concentration ◽  
Successful Attempt ◽  
Cell Assays ◽  
Mixing Method

Abstract In recent years, the potential of nanobubbles (NBs) for biological activation has been actively investigated. In this study, we investigated the proliferative effects of nitrogen nanobubbles (N-NBs) on fibroblast cells using cell assays with image analysis and flow cytometry. A high concentration of N-NBs (more than 4 × 108 NBs/mL) was generated in Dulbecco’s modified Eagle’s medium (DMEM) using a gas-liquid mixing method. In image analysis, the cells were counted and compared, which showed an 11% improvement in proliferation in the culture medium with N-NBs. In flow cytometry, the decrease in the fluorescence intensity was analyzed, which revealed a 1.5% improvement in proliferation in the culture medium with N-NBs. This study represents the first successful attempt of directly generating quantified NBs in a culture medium for cell culture. The findings suggest that the N-NBs in the culture medium can facilitate cell proliferation.

scholarly journals 3D-Printed Bioreactor with Integrated Impedance Spectroscopy for Cell Barrier Monitoring

2021 ◽  
Author(s):  
Matthias Wessling
Keyword(s):  
Cell Culture ◽  
Epithelial Cell ◽  
Impedance Spectroscopy ◽  
Indium Tin Oxide ◽  
Electric Circuit ◽  
Electrical Impedance Spectroscopy ◽  
Oxide Glass ◽  
Epithelial Cell Line ◽  
Scientific Institutions ◽  
3D Printed

Cell culture experiments often suffer from limited commercial availability of laboratory-scale bioreactors, which allow experiments to be conducted under flow conditions and additional online monitoring techniques. A novel 3D-printed bioreactor with a homogeneously distributed flow field enabling epithelial cell culture experiments and online barrier monitoring by integrated electrodes through electrical impedance spectroscopy (EIS) is presented. Transparent and conductive indium tin oxide glass as current-injecting electrodes allows direct visualization of the cells, while measuring EIS simultaneously. The bioreactor's design considers the importance of a homogeneous electric field by placing the voltage pick-up electrodes in the electrical field. The device's functionality is demonstrated by the cultivation of the epithelial cell line Caco-2 under continuous flow and monitoring of the cell layer by online EIS. The collected EIS data were fitted by an equivalent electric circuit, resulting in the cell layer's resistance and capacitance. This data is used to monitor the cell layer's reaction to ethylene glycol-bis-(2-aminoethyl ether)-N,N,N′,N′-tetraacetic acid and forskolin. These two model substances show the power of impedance spectroscopy as a non-invasive way to characterize cell barriers. In addition, the bioreactor design is available as a print-ready file in the Appendix, enabling its use for other scientific institutions.

scholarly journals Dielectrophoretic and Electrical Impedance Differentiation of Cancerous Cells Based on Biophysical Phenotype

Biosensors ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 401
Author(s):  
Ina Turcan ◽  
Iuliana Caras ◽  
Thomas Gabriel Schreiner ◽  
Catalin Tucureanu ◽  
Aurora Salageanu ◽  
...  
Keyword(s):  
Impedance Spectroscopy ◽  
Tumor Cells ◽  
Cancer Cell ◽  
Primary Tumor ◽  
Electrical Impedance ◽  
Mononuclear Cells ◽  
Equivalent Circuit Model ◽  
Electrical Impedance Spectroscopy ◽  
Label Free ◽  
Peripheral Blood Mononuclear

Here, we reported a study on the detection and electrical characterization of both cancer cell line and primary tumor cells. Dielectrophoresis (DEP) and electrical impedance spectroscopy (EIS) were jointly employed to enable the rapid and label-free differentiation of various cancer cells from normal ones. The primary tumor cells that were collected from two colorectal cancer patients and cancer cell lines (SW-403, Jurkat, and THP-1), and healthy peripheral blood mononuclear cells (PBMCs) were trapped first at the level of interdigitated microelectrodes with the help of dielectrophoresis. Correlation of the cells dielectric characteristics that was obtained via electrical impedance spectroscopy (EIS) allowed evident differentiation of the various types of cell. The differentiations were assigned to a “dielectric phenotype” based on their crossover frequencies. Finally, Randles equivalent circuit model was employed for highlighting the differences with regard to a series group of charge transport resistance and constant phase element for cancerous and normal cells.

scholarly journals EVALUATION OF MICROMOTION OF VASCULAR ENDOTHELIAL CELLS IN ELECTRICAL CELL-SUBSTRATE IMPEDANCE SENSING (ECIS) METHOD USING A MATHEMATICAL MODEL

2005 ◽  
Vol 05 (02) ◽  
pp. 357-368 ◽  
Author(s):  
NORIKO GODA ◽  
NORIYUKI KATAOKA ◽  
JUICHIRO SHIMIZU ◽  
SATOSHI MOHRI ◽  
YOSHITAKE YAMAMOTO ◽  
...  
Keyword(s):  
Mathematical Model ◽  
Endothelial Cells ◽  
Cell Membrane ◽  
Culture Medium ◽  
Cultured Cells ◽  
Vascular Endothelial Cells ◽  
Shielding Effect ◽  
Total Resistance ◽  
Impedance Sensing ◽  
Cell Substrate

We proposed a mathematical model for the microdynamics of cultured cells measured with ECIS (Electrical Cell-substrate Impedance Sensing) system that can separately evaluate cell-to-cell and cell-to-substrate gaps. Our mathematical model is composed of culture medium impedance between cells (Zsol), cell impedance (Zc), and polarization impedance of the electrode (Zp). Zsol consists of the resistance between cells (Rsol) and the capacitance between cells (Csol) of the culture medium. In particular, Rsol is the resistance component related to the cell-to-cell distance (A). Zc consists of capacitance of the cell membrane (Cc) and resistance of the cell membrane (Rc). Zp depends on the cell-to-substrate distance (h) because of the shielding effect of cells to the electrode. The shielding effect is defined as shielding coefficient (Sk). We examined the changes in the impedance of the electrode without or with cells in various conditions. The electrical characteristics of the electrode with or without cells agreed well with those measured in ECIS system. It was found that whether A or h caused the changes in the impedance could be determined based on the changes in the total resistance and reactance (capacitance); A mainly affects the total resistance value, and h mainly affects total capacitance value. Therefore, we can simply estimate the changes in cell-to-cell and cell-to-substrate gaps with measured total resistance and reactance (capacitance). Based on these results, when the cultured endothelial cells (HUVEC) were stimulated with estrogen for 40 hours, it was proved that the cell-to-cell distances decreased, even though the cell-to-electrode distances slightly increased. This result suggests that the barrier function of endothelium is fortified by estrogen.

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