scholarly journals In Vitro Wounding Models Using the Electric Cell-Substrate Impedance Sensing (ECIS)-Zθ Technology

Biosensors ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 90 ◽  
Author(s):  
Andrea Gu ◽  
Dan Kho ◽  
Rebecca Johnson ◽  
E. Graham ◽  
Simon O’Carroll
Keyword(s):  
Barrier Function ◽  
Cell Monolayer ◽  
Immunohistochemical Analysis ◽  
Cell Loss ◽  
Impedance Sensing ◽  
Viable Cells ◽  
Cell Substrate ◽  
Underlying Causes ◽  
A Cell

Electric Cell-Substrate Impedance Sensing (ECIS) can produce reproducible wounding models by mechanically disrupting a cell monolayer. This study compared in vitro wound-healing using human cerebral microvascular endothelial cells (hCMVEC) with both single electrode (8W1E) and multiple electrodes (8W10E+) arrays. Measurements of hCMVEC migration and barrier functions were conducted, revealing variable levels of barrier disruption could be achieved by altering the duration and magnitude of the applied current. In all scenarios, the barrier (Rb) did not recover the strength observed prior to injury. Localization of junctional proteins following wounding were analyzed by immunocytochemistry. Following wounding, cell migration was generally faster on the 8W10E+ than the 8W1E array. Immunohistochemical analysis revealed non-viable cells remained on the 8W1E electrodes but not the 8W10E+ electrodes. However, viable cells partially remained on the 8W10E+ electrodes following wounding. In addition, the 8W10E+ electrodes demonstrated variation in cell loss across electrodes within the same well. This suggests the type of wounding is different on the two array types. However, our data show both arrays can be used to model incomplete barrier recovery and therefore both have potential for testing of drugs to improve endothelial barrier function. This is the first time that the possibility of using the 8W10E+ array as a wounding model is addressed. We highlight the differences in wounding produced between the two arrays, and can be used to study the underlying causes for impaired barrier function following CNS injuries.

Cell-Based Detection Using Electric Cell-Impedance Sensing

2009 ◽  
Vol 1239 ◽  
Author(s):  
Bhavana Mohanraj ◽  
Nathan Schiele ◽  
Anne Hynes ◽  
Zijie Yan ◽  
David T. Corr ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Barrier Function ◽  
Cell Monolayer ◽  
Significant Drop ◽  
Impedance Sensing ◽  
Cell Impedance ◽  
A Cell ◽  
Measured Resistance ◽  
20 Nm ◽  
Future Work

AbstractElectric Cell-Impedance Sensing (ECIS) is a real-time transduction system that can be used to detect the presence of foreign particles or pathogens by measuring the changes in impedance or resistance of a cell monolayer grown on an electrode. Herein, we present the use of ECIS for the detection of the toxicity of silver nanoparticles on Madine Derby Canine Kidney (MDCK) epithelial cells as a function of changes in the cell confluence and barrier function of the cell monolayer. The barrier function is a measure of the number of tight junctions formed between confluent cells in a monolayer; tighter confluence leads to an increase in a barrier function and thus in the measured resistance. We were able to detect exposures as low as 1 μg of 20 nm silver nanoparticles per 105 cells within 2 hours; those exposures were quantified as a significant drop in impedance and a gradual decrease in the barrier function as compared to the controls. Future work would include the detection of protein toxins using impedance sensing as well as further analysis of the barrier function using fluorescent staining.

Analysis of the Dynamics of the Electric Capacitance of a Cell Monolayer at the Late Stages of Caco-2 cell Differentiation

2019 ◽  
Vol 35 (6) ◽  
pp. 87-90
Author(s):  
S.V. Nikulin ◽  
V.A. Petrov ◽  
D.A. Sakharov
Keyword(s):  
Impedance Spectroscopy ◽  
Cell Monolayer ◽  
Microfluidic Chips ◽  
Russian Science ◽  
Electric Capacitance ◽  
A Cell ◽  
Static Conditions ◽  
Late Stages

The real-time monitoring of electric capacitance (impedance spectroscopy) allowed obtaining evidence that structures which look like intestinal villi can be formed during the cultivation under static conditions as well as during the cultivation in microfluidic chips. It was shown in this work via transcriptome analysis that the Hh signaling pathway is involved in the formation of villus-like structures in vitro, which was previously shown for their formation in vivo. impedance spectroscopy, intestine, villi, electric capacitance, Hh The study was funded by the Russian Science Foundation (Project 16-19-10597).

Manipulating the Barrier Function of a Cell Monolayer Using a High-power Miniature Ultrasonic Transducer

2021 ◽  
Author(s):  
Mihnea V. Turcanu ◽  
Alexandru C. Moldovan ◽  
Maya Thanou ◽  
Inke Nathke ◽  
Sandy Cochran
Keyword(s):  
High Power ◽  
Barrier Function ◽  
Ultrasonic Transducer ◽  
Cell Monolayer ◽  
A Cell

scholarly journals Ethanol-Induced Lymphatic Endothelial Cell Permeability via MAP-Kinase Regulation

2021 ◽  
Author(s):  
Matthew Herrera ◽  
Patricia Molina ◽  
Flavia M. Souza-Smith
Keyword(s):  
Endothelial Cell ◽  
Cell Viability ◽  
Barrier Function ◽  
Protein Localization ◽  
Cell Monolayer ◽  
Lymphatic Endothelial Cell ◽  
Cell Permeability ◽  
Gene Expressions ◽  
Mapk Activation

Chronic alcohol alters the immune system enhancing the susceptibility to inflammation, bacterial, and viral infections in alcohol users. We have shown that alcohol causes increased permeability of mesenteric lymphatic vessels in alcohol fed rats. The mechanisms of alcohol-induced lymphatic leakage are unknown. Endothelial cell monolayer permeability is controlled by junctional proteins complexes called tight junctions (TJ) and adherens junctions (AJ), and each can be regulated by MAPK activation. We hypothesize that ethanol induces lymphatic endothelial cell (LEC) permeability via disruption of LEC TJ through MAPK activation. An in vitro model of rat LECs was used. Ethanol-supplemented medium was added at concentrations of 0, 25, and 50 mM to confluent cells. Resistance-based barrier function, transwell permeability, cell viability, TJ, AJ, and MAPK protein activity, TJ and AJ gene expressions, and the role of p38 MAPK in barrier function regulation were measured. Ethanol increased the permeability of LECs compared to controls that was not associated with decreased cell viability. LECs treated with 50 mM ethanol showed an increase in phosphorylated levels of p38. No significant changes in TJ and AJ gene or protein expressions were observed after ethanol treatment. p38 inhibition prevented ethanol-induced increases in permeability. These findings suggest that p38 may play a role in the regulation of ethanol-induced LEC permeability, but altered permeability may not be associated with decreased TJ or AJ protein expression. Further investigation into junctional protein localization is needed to better understand the effects of ethanol on lymphatic endothelial cell-to-cell contacts and hyperpermeability.

scholarly journals 28 CLONED EMBRYOS CAN BE PRODUCED USING DONOR CELLS OBTAINED FROM A 72-HOUR COOLED CARCASS

2005 ◽  
Vol 17 (2) ◽  
pp. 164 ◽  
Author(s):  
S. Arat ◽  
H. Bagis ◽  
H. Odaman Mercan ◽  
A. Dinnyes
Keyword(s):  
Calf Serum ◽  
Calcium Ionophore ◽  
Cumulus Cells ◽  
Penicillin G ◽  
Developmental Potential ◽  
Viable Cells ◽  
Donor Cells ◽  
A Cell ◽  
Vitro Development

There are few reports on the use of cells from a dead mammal for nuclear transfer (NT). So far, most calves have been cloned from live adult cows or fresh fetal samples. The ability to produce cloned animals using postmortem tissue can provide an additional application to the field of NT. This study was conducted to investigate whether viable cells could be obtained from tissues chilled for 72 h and whether these cells could be used for NT. Bovine oocytes isolated from slaughterhouse ovaries were matured in TCM199 supplemented with 10% fetal calf serum (FBS), 50 μg/mL sodium pyruvate, 1% v:v penicillin-streptomycin (10,000 U/mL penicillin G, 10,000 μg/mL streptomycin), 10 ng/mL EGF, 0.5 μg/mL FSH, and 5 μg/mL LH. A cell line (MC) was established from leg muscle of a cow carcass stored at 0°C for 72 h. Tissues from muscle were cut into small pieces. Tissue explants were cultured in DMEM-F12 supplemented with 10% FBS at 37°C in 5% CO2 in air. Bovine granulosa cells (GC) were isolated from ovarian follicles and used for NT as control cells. Prior to NT, all somatic cells were allowed to grow to confluency (G1/G0) in DMEM-F12 medium supplemented with 10% FBS. Cumulus cells were removed by vortexing with hyaluronidase at 18 h after the start of maturation. Matured oocytes labeled with DNA fluorochrome Hoechst 33342 were enucleated under UV to ensure full removal of the chromatin. A single cell was inserted into the perivitelline space of the enucleated oocyte. Oocyte-cell couples were fused by a DC pulse of 133V/500 μm for 25 μs. After fusion, NT units were activated using a combination of calcium ionophore (5 μM), cytochalasin D (2.5 μg/mL) and cycloheximide (10 μg/mL) and cultured for 7 days in BARC or G1.3-G2.3 medium. Differences (developmental potential and cell numbers) among groups were analyzed by one-way ANOVA after arcsin square transformation. The results are summarized in Table 1. The results suggest that viable cells can be obtained from muscle of a cow carcass stored at cold temperature for 72 h and that these cells have ability to generate NT blastocysts at rates similar to those obtained with fresh GCs. In addition, G1.3 and G2.3 culture medium supported embryo development better than BARC medium. Table 1. In vitro development of NT embryos This study was supported by a grant from TUBITAK, Turkey (VHAG-1908 and Turkey-Hungary bilateral project VHAG-2022).

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