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 Cell Barrier Characterization in Transwell Inserts by Electrical Impedance Spectroscopy

2020 ◽  
Author(s):  
Matthias Wessling
Keyword(s):  
Impedance Spectroscopy ◽  
Electrical Impedance ◽  
Cell Layer ◽  
Electric Circuit ◽  
Electrical Impedance Spectroscopy ◽  
Coating Materials ◽  
Simple Method ◽  
Capacitive Properties ◽  
Membrane Coating ◽  
Permeability Enhancers

We describe an impedance-based method for cell barrier integrity testing. A four-electrode electrical impedance spectroscopy (EIS) setup can be realized by simply connecting a commercial chopstick-like electrode (STX-1) to a potentiostat allowing monitoring cell barriers cultivated in transwell inserts.Subsequent electric circuit modeling of the electrical impedance results the capacitive properties of the barrier next to the well-known transepithelial electrical resistance (TEER). The versatility of the newmethod was analyzed by the EIS analysis of a Caco-2 monolayer in response to (a) different membrane coating materials, (b) two different permeability enhancers ethylene glycol-bis(2-aminoethylether)-N,N,N’,N’-tetraacetic acid (EGTA) and saponin, and (c) sonoporation. For the different membrane coating materials, the TEERs of the standard and new protocol coincide and increase during cultivation, while the capacitance shows a distinct maximum for three different surface materials (no coating, Matrigel®, and collagen I). The permeability enhancers cause a decline in the TEER value, but only saponin alters the capacitance of the cell layer by two orders of magnitude. Hence, cell layer capacitance and TEER represent two independent properties characterizing the monolayer. The use of commercial chopstick-like electrodes to access the impedance of a barrier cultivated in transwell inserts enables remarkable insight into the behavior of the cellular barrier with no extra work for the researcher. This simple method could evolve into a standard protocol used in cell barrier research.

scholarly journals 3D-Printed Graphene Electrodes Applied in an Impedimetric Electronic Tongue for Soil Analysis

Chemosensors ◽  
2019 ◽  
Vol 7 (4) ◽  
pp. 50 ◽  
Author(s):  
Americo da Silva ◽  
Braunger ◽  
Neris Coutinho ◽  
Rios do Amaral ◽  
Rodrigues ◽  
...  
Keyword(s):  
Impedance Spectroscopy ◽  
Precision Agriculture ◽  
Soil Analysis ◽  
Electronic Tongue ◽  
Soil Samples ◽  
Electrical Impedance Spectroscopy ◽  
World Population ◽  
Fused Deposition ◽  
Lbl Films ◽  
3D Printed

The increasing world population leads to the growing demand for food production without expanding cultivation areas. In this sense, precision agriculture optimizes the production and input usage by employing sensors to locally monitor plant nutrient within agricultural fields. Here, we have used an electronic tongue sensing device based on impedance spectroscopy to recognize distinct soil samples (sandy and clayey) enriched with macronutrients. The e-tongue setup consisted of an array of four sensing units formed by layer-by-layer (LbL) films deposited onto 3D-printed graphene-based interdigitated electrodes (IDEs). The IDEs were fabricated in 20 min using the fused deposition modeling process and commercial polylactic acid-based graphene filaments. The e-tongue comprised one bare and three IDEs functionalized with poly(diallyldimethylammonium chloride) solution/copper phthalocyanine-3,4′,4′′,4′′′-tetrasulfonic acid tetrasodium salt (PDDA/CuTsPc), PDDA/montmorillonite clay (MMt-K), and PDDA/poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) LbL films. Control samples of sandy and clayey soils were enriched with different concentrations of nitrogen (N), phosphorus (P), and potassium (K) macronutrients. Sixteen soil samples were simply diluted in water and measured using electrical impedance spectroscopy, with data analyzed by principal component analysis. All soil samples were easily distinguished without pre-treatment, indicating the suitability of 3D-printed electrodes in e-tongue analysis to distinguish the chemical fertility of soil samples. Our results encourage further investigations into the development of new tools to support precision agriculture.

Development of the microsporidian parasite, Loma salmonae, in a rainbow trout gill epithelial cell line (RTG-1): evidence of xenoma development in vitro

Parasitology ◽  
2014 ◽  
Vol 142 (2) ◽  
pp. 326-331 ◽  
Author(s):  
S. H. McCONNACHIE ◽  
J. SHEPPARD ◽  
G. M. WRIGHT ◽  
D. J. SPEARE
Keyword(s):  
Cell Culture ◽  
Rainbow Trout ◽  
Epithelial Cell ◽  
Cell Line ◽  
Cell Model ◽  
Epithelial Cell Line ◽  
Microsporidian Parasite ◽  
Loma Salmonae ◽  
Trout Gill

SUMMARYGrowth and propagation of fish-infecting microsporidians within cell culture has been more difficult to achieve than for insect- and human-infecting microsporidians. Fish microsporidia tend to elicit xenoma development rather than diffuse growth in vivo, and this process likely increases host specificity. We present evidence that the fish microsporidian, Loma salmonae, has the capacity to develop xenomas within a rainbow trout gill epithelial cell line (RTG-1). Spore numbers increased over a 4 weeks period within cell culture flasks. Xenoma-like structures were observed using phase contrast microscopy, and then confirmed using transmission electron microscopy. Optimization of the L. salmonae-RTG-1 cell model has important implications in elucidating the process of xenoma development induced by microsporidian parasites.

scholarly journals Suramin inhibits SARS-CoV-2 infection in cell culture by interfering with early steps of the replication cycle

2020 ◽  
Author(s):  
Clarisse Salgado Benvindo da Silva ◽  
Melissa Thaler ◽  
Ali Tas ◽  
Natacha S. Ogando ◽  
Peter J. Bredenbeek ◽  
...  
Keyword(s):  
Cell Culture ◽  
Epithelial Cell ◽  
Randomized Clinical Trials ◽  
Drug Repurposing ◽  
Preclinical Study ◽  
Replication Cycle ◽  
Epithelial Cell Line ◽  
Airway Epithelial ◽  
Daily Lives ◽  
Human Lung Epithelial Cell

AbstractThe SARS-CoV-2 pandemic that originated from Wuhan, China, in December 2019 has impacted public health, society and economy and the daily lives of billions of people in an unprecedented manner. There are currently no specific registered antiviral drugs to treat or prevent SARS-CoV-2 infections. Therefore, drug repurposing would be the fastest route to provide at least a temporary solution while better, more specific drugs are being developed. Here we demonstrate that the antiparasitic drug suramin inhibits SARS-CoV-2 replication, protecting Vero E6 cells with an EC50 of ∼20 µM, which is well below the maximum attainable level in human serum. Suramin also decreased the viral load by 2-3 logs when Vero E6 cells or cells of a human lung epithelial cell line (Calu-3) were treated. Time of addition and plaque reduction assays showed that suramin acts on early steps of the replication cycle, possibly preventing entry of the virus. In a primary human airway epithelial cell culture model, suramin also inhibited the progression of infection. The results of our preclinical study warrant further investigation and suggest it is worth evaluating whether suramin provides any benefit for COVID-19 patients, which obviously requires well-designed, properly controlled randomized clinical trials.

scholarly journals 3D-Printed Sensors and Actuators in Cell Culture and Tissue Engineering: Framework and Research Challenges

Sensors ◽  
2020 ◽  
Vol 20 (19) ◽  
pp. 5617
Author(s):  
Pablo Pérez ◽  
Juan Alfonso Serrano ◽  
Alberto Olmo
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
3D Printing ◽  
Regenerative Medicine ◽  
Electrical Impedance Spectroscopy ◽  
Special Focus ◽  
Tissue Formation ◽  
Sensors And Actuators ◽  
Printed Sensors ◽  
3D Printed

Three-dimensional printing technologies have been recently proposed to monitor cell cultures and implement cell bioreactors for different biological applications. In tissue engineering, the control of tissue formation is crucial to form tissue constructs of clinical relevance, and 3D printing technologies can also play an important role for this purpose. In this work, we study 3D-printed sensors that have been recently used in cell culture and tissue engineering applications in biological laboratories, with a special focus on the technique of electrical impedance spectroscopy. Furthermore, we study new 3D-printed actuators used for the stimulation of stem cells cultures, which is of high importance in the process of tissue formation and regenerative medicine. Key challenges and open issues, such as the use of 3D printing techniques in implantable devices for regenerative medicine, are also discussed.

scholarly journals Suramin Inhibits SARS-CoV-2 Infection in Cell Culture by Interfering with Early Steps of the Replication Cycle

2020 ◽  
Vol 64 (8) ◽  
Author(s):  
Clarisse Salgado-Benvindo ◽  
Melissa Thaler ◽  
Ali Tas ◽  
Natacha S. Ogando ◽  
Peter J. Bredenbeek ◽  
...  
Keyword(s):  
Cell Culture ◽  
Epithelial Cell ◽  
Global Economy ◽  
Randomized Clinical Trials ◽  
Drug Repurposing ◽  
Preclinical Study ◽  
Replication Cycle ◽  
Epithelial Cell Line ◽  
Airway Epithelial ◽  
Human Lung Epithelial Cell

ABSTRACT The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic that originated in Wuhan, China, in December 2019 has impacted public health, society, the global economy, and the daily lives of billions of people in an unprecedented manner. There are currently no specific registered antiviral drugs to treat or prevent SARS-CoV-2 infections. Therefore, drug repurposing would be the fastest route to provide at least a temporary solution while better, more specific drugs are being developed. Here, we demonstrate that the antiparasitic drug suramin inhibits SARS-CoV-2 replication, protecting Vero E6 cells with a 50% effective concentration (EC50) of ∼20 μM, which is well below the maximum attainable level in human serum. Suramin also decreased the viral load by 2 to 3 logs when Vero E6 cells or cells of a human lung epithelial cell line (Calu-3 2B4 [referred to here as “Calu-3”]) were treated. Time-of-addition and plaque reduction assays performed on Vero E6 cells showed that suramin acts on early steps of the replication cycle, possibly preventing binding or entry of the virus. In a primary human airway epithelial cell culture model, suramin also inhibited the progression of infection. The results of our preclinical study warrant further investigation and suggest that it is worth evaluating whether suramin provides any benefit for COVID-19 patients, which obviously requires safety studies and well-designed, properly controlled randomized clinical trials.

Sign in / Sign up

Export Citation Format

Share Document