scholarly journals Real-time quantitative monitoring of hiPSC-based model of macular degeneration on Electric Cell-substrate Impedance Sensing microelectrodes

2015 ◽  
Vol 71 ◽  
pp. 445-455 ◽  
Author(s):  
W. Gamal ◽  
S. Borooah ◽  
S. Smith ◽  
I. Underwood ◽  
V. Srsen ◽  
...  
Keyword(s):  
Macular Degeneration ◽  
Real Time ◽  
Impedance Sensing ◽  
Cell Substrate ◽  
Quantitative Monitoring

scholarly journals Electric Cell-Substrate Impedance Sensing To Monitor Viral Growth and Study Cellular Responses to Infection with Alphaherpesviruses in Real Time

mSphere ◽  
2017 ◽  
Vol 2 (2) ◽  
Author(s):  
Matthew R. Pennington ◽  
Gerlinde R. Van de Walle
Keyword(s):  
Real Time ◽  
Cellular Response ◽  
Cellular Responses ◽  
Cellular Damage ◽  
Impedance Sensing ◽  
Novel Technologies ◽  
Antiviral Therapies ◽  
Mucosal Surfaces ◽  
Cell Substrate ◽  
Hsv 1

ABSTRACT Alphaherpesviruses, including those that commonly infect humans, such as HSV-1 and HSV-2, typically infect and cause cellular damage to epithelial cells at mucosal surfaces, leading to disease. The development of novel technologies to study the cellular responses to infection may allow a more complete understanding of virus replication and the creation of novel antiviral therapies. This study demonstrates the use of ECIS to study various aspects of herpesvirus biology, with a specific focus on changes in cellular morphology as a result of infection. We conclude that ECIS represents a valuable new tool with which to study alphaherpesvirus infections in real time and in an objective and reproducible manner. Electric cell-substrate impedance sensing (ECIS) measures changes in an electrical circuit formed in a culture dish. As cells grow over a gold electrode, they block the flow of electricity and this is read as an increase in electrical impedance in the circuit. ECIS has previously been used in a variety of applications to study cell growth, migration, and behavior in response to stimuli in real time and without the need for cellular labels. Here, we demonstrate that ECIS is also a valuable tool with which to study infection by alphaherpesviruses. To this end, we used ECIS to study the kinetics of cells infected with felid herpesvirus type 1 (FHV-1), a close relative of the human alphaherpesviruses herpes simplex virus 1 (HSV-1) and HSV-2, and compared the results to those obtained with conventional infectivity assays. First, we demonstrated that ECIS can easily distinguish between wells of cells infected with different amounts of FHV-1 and provides information about the cellular response to infection. Second, we found ECIS useful in identifying differences between the replication kinetics of recombinant DsRed Express2-labeled FHV-1, created via CRISPR/Cas9 genome engineering, and wild-type FHV-1. Finally, we demonstrated that ECIS can accurately determine the half-maximal effective concentration of antivirals. Collectively, our data show that ECIS, in conjunction with current methodologies, is a powerful tool that can be used to monitor viral growth and study the cellular response to alphaherpesvirus infection. IMPORTANCE Alphaherpesviruses, including those that commonly infect humans, such as HSV-1 and HSV-2, typically infect and cause cellular damage to epithelial cells at mucosal surfaces, leading to disease. The development of novel technologies to study the cellular responses to infection may allow a more complete understanding of virus replication and the creation of novel antiviral therapies. This study demonstrates the use of ECIS to study various aspects of herpesvirus biology, with a specific focus on changes in cellular morphology as a result of infection. We conclude that ECIS represents a valuable new tool with which to study alphaherpesvirus infections in real time and in an objective and reproducible manner.

scholarly journals Comparison of Leading Biosensor Technologies to Detect Changes in Human Endothelial Barrier Properties in Response to Pro-Inflammatory TNFα and IL1β in Real-Time

Biosensors ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 159
Author(s):  
James J. W. Hucklesby ◽  
Akshata Anchan ◽  
Simon J. O'Carroll ◽  
Charles P. Unsworth ◽  
E. Scott Graham ◽  
...  
Keyword(s):  
Human Brain ◽  
Real Time ◽  
Barrier Properties ◽  
Endothelial Barrier ◽  
Endothelial Monolayer ◽  
Impedance Sensing ◽  
Endothelial Monolayers ◽  
Impedance Data ◽  
Cell Substrate ◽  
Limited Frequency

Electric Cell-Substrate Impedance Sensing (ECIS), xCELLigence and cellZscope are commercially available instruments that measure the impedance of cellular monolayers. Despite widespread use of these systems individually, direct comparisons between these platforms have not been published. To compare these instruments, the responses of human brain endothelial monolayers to TNFα and IL1β were measured on all three platforms simultaneously. All instruments detected transient changes in impedance in response to the cytokines, although the response magnitude varied, with ECIS being the most sensitive. ECIS and cellZscope were also able to attribute responses to particular endothelial barrier components by modelling the multifrequency impedance data acquired by these instruments; in contrast the limited frequency xCELLigence data cannot be modelled. Consistent with its superior impedance sensing, ECIS exhibited a greater capacity than cellZscope to distinguish between subtle changes in modelled endothelial monolayer properties. The reduced resolving ability of the cellZscope platform may be due to its electrode configuration, which is necessary to allow access to the basolateral compartment, an important advantage of this instrument. Collectively, this work demonstrates that instruments must be carefully selected to ensure they are appropriate for the experimental questions being asked when assessing endothelial barrier properties.

scholarly journals Real-Time Measurement of Melanoma Cell-Mediated Human Brain Endothelial Barrier Disruption Using Electric Cell-Substrate Impedance Sensing Technology

Biosensors ◽  
2019 ◽  
Vol 9 (2) ◽  
pp. 56 ◽  
Author(s):  
Akshata Anchan ◽  
Panagiota Kalogirou-Baldwin ◽  
Rebecca Johnson ◽  
Dan T Kho ◽  
Wayne Joseph ◽  
...  
Keyword(s):  
Human Brain ◽  
Real Time ◽  
Melanoma Cell ◽  
Time Lapse ◽  
Melanoma Cells ◽  
Human Melanoma ◽  
Brain Endothelium ◽  
Impedance Sensing ◽  
Sensing Technology ◽  
Cell Substrate

Electric cell-substrate impedance sensing (ECIS) is an impedance-based method for monitoring changes in cell behaviour in real-time. In this paper, we highlight the importance of ECIS in measuring the kinetics of human melanoma cell invasion across human brain endothelium. ECIS data can be mathematically modelled to assess which component of the endothelial paracellular and basolateral barriers is being affected and when. Our results reveal that a range of human melanoma cells can mediate disruption of human brain endothelium, primarily involving the paracellular route, as demonstrated by ECIS. The sensitivity of ECIS also reveals that the paracellular barrier weakens within 30–60 min of the melanoma cells being added to the apical face of the endothelial cells. Imaging reveals pronounced localisation of the melanoma cells at the paracellular junctions consistent with paracellular migration. Time-lapse imaging further reveals junctional opening and disruption of the endothelial monolayer by the invasive melanoma cells all within several hours. We suggest that the ability of ECIS to resolve changes to barrier integrity in real time, and to determine the route of migration, provides a powerful tool for future studies investigating the key molecules involved in the invasive process of cancer cells.

scholarly journals Real-Time Monitoring of Neural Differentiation of Human Mesenchymal Stem Cells by Electric Cell-Substrate Impedance Sensing

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Hyo Eun Park ◽  
Donghee Kim ◽  
Hyun Sook Koh ◽  
Sungbo Cho ◽  
Jung-Suk Sung ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Real Time ◽  
Neural Differentiation ◽  
Human Mesenchymal Stem Cells ◽  
Real Time Monitoring ◽  
Impedance Sensing ◽  
Cell Substrate ◽  
Cells Cultured

Stem cells are useful for cell replacement therapy. Stem cell differentiation must be monitored thoroughly and precisely prior to transplantation. In this study we evaluated the usefulness of electric cell-substrate impedance sensing (ECIS) forin vitroreal-time monitoring of neural differentiation of human mesenchymal stem cells (hMSCs). We cultured hMSCs in neural differentiation media (NDM) for 6 days and examined the time-course of impedance changes with an ECIS array. We also monitored the expression of markers for neural differentiation, total cell count, and cell cycle profiles. Cellular expression of neuron and oligodendrocyte markers increased. The resistance value of cells cultured in NDM was automatically measured in real-time and found to increase much more slowly over time compared to cells cultured in non-differentiation media. The relatively slow resistance changes observed in differentiating MSCs were determined to be due to their lower growth capacity achieved by induction of cell cycle arrest in G0/G1. Overall results suggest that the relatively slow change in resistance values measured by ECIS method can be used as a parameter for slowly growing neural-differentiating cells. However, to enhance the competence of ECIS forin vitroreal-time monitoring of neural differentiation of MSCs, more elaborate studies are needed.

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