The development of an epithelial barrier was assessed via TEER measurements, recorded with ERS-2 Volt-Ohm-Meter with STX01 chopstick electrodes (Millipore, Burlington, MA, USA) on each day of submerged culture and on distinct days of ALI culture. Medium was replaced in both Transwell compartments and cells were incubated at 37 °C for 25 min and subsequently kept at room temperature for 2 min before measuring TEER. Values were recorded for each of the three cavities of one insert and the mean was calculated. The values of a blank insert without cells were subtracted as the background value.
Stx01 chopstick electrode
Manufactured by Merck Group
Sourced in United States
The STX01 chopstick electrode is a laboratory instrument used for electrochemical measurements. It features two parallel electrodes in a chopstick-like design. The core function of this product is to facilitate precise and controlled electrochemical experiments and analyses.
Automatically generated - may contain errors
Lab products found in correlation
Millicell ers 2, by Merck Group (3 mentions)
Millicell ers 2 voltohmmeter, by Merck Group (2 mentions)
Ers 2 voltohmmeter, by Merck Group (1 mentions)
Prism 5, by GraphPad (1 mentions)
Ers 2 epithelial volt ohm meter, by Merck Group (1 mentions)
Millicell ers 2 epithelial volt ohm meter, by Merck Group (1 mentions)
8 protocols using stx01 chopstick electrode
1
Assessing Epithelial Barrier Development
2
Evaluating In Vitro Blood-Brain Barrier Integrity
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TEER was measured to evaluate the integrity of the in vitro BBB models. TEER, which in culture conditions reflects the flux of mainly sodium ions through an intact cell layer, was measured using a Millicell ERS-2 epithelial Volt-Ohm meter and STX01 Chopstick Electrodes (Millipore, Hellerup Denmark, DK). The TEER values of coated but cell-free inserts were subtracted from the measured TEER values, and the difference was multiplied with the size of the insert (1.12 cm2). Measured TEER values are given as Ω cm2. Data were analysed with GraphPad Prism 5.0 software (GraphPad Software, Inc., CA, USA) using two-way ANOVA with Bonferroni post hoc test.
3
In Vitro Co-culture Model of the Blood-Brain Barrier
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To construct in vitro co-culture models of the BBB, BCECs were detached from the culture dish and seeded onto the porous membrane of hanging cell culture inserts fitted for 12-well plates (Transwell, polyethylene terephthalate porous membrane, pore size: 1 µm). All culture inserts were coated with collagen IV and fibronectin. To increase the tightness of the BCECs, the hanging cell culture inserts were transferred to a 12-well plate containing a confluent layer of mixed glial cells (mostly astrocytes) in the bottom of each well, and treated with hydrocortisone, cAMP and RO-201724 at concentrations of 550 nM, 250 μM and 17.5 μM, respectively.
The integrity of the in vitro BBB was evaluated by measuring the transendothelial electrical resistance (TEER) using the Millicell ERS-2 Epithelial Volt-Ohm meter and STX01 Chopstick Electrodes (Millipore, Hellerup, DK). The corresponding permeability parameters to the TEER values of the in vitro BBB models were as presented in Thomsen et al. 26 (link). The TEER value of a coated insert without any cells were subtracted from all TEER measurements as normalization, and all values multiplied by the area of the culture insert (1.12 cm2). All data were analyzed using GraphPad 5.0 (GraphPad Software, Inc., CA, USA), and the TEER values presented as Ω cm2.
The integrity of the in vitro BBB was evaluated by measuring the transendothelial electrical resistance (TEER) using the Millicell ERS-2 Epithelial Volt-Ohm meter and STX01 Chopstick Electrodes (Millipore, Hellerup, DK). The corresponding permeability parameters to the TEER values of the in vitro BBB models were as presented in Thomsen et al. 26 (link). The TEER value of a coated insert without any cells were subtracted from all TEER measurements as normalization, and all values multiplied by the area of the culture insert (1.12 cm2). All data were analyzed using GraphPad 5.0 (GraphPad Software, Inc., CA, USA), and the TEER values presented as Ω cm2.
4
Evaluating Cellular Barrier Integrity with TEER
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The electrical parameter, trans-endothelial electrical resistance (TEER), was utilized to evaluate the integrity of the cellular barriers in vitro following exposure to CS. TEER was measured using a Millicell-ERS-2 (Millipore, Billerica, MA, US) volt-ohm-meter equipped with a STX01 chopstick electrode (Millipore, Billerica, MA, US). Three replicate measurements were taken for each well, and three wells were tested per experimental run. The obtained values were adjusted with air controls and then multiplied by the effective membrane area in cm2 (1.12 cm2 for 12-well transwell inserts) to obtain the final result in Ω cm2. TEER measurements allowed for the assessment of cell layer integrity and toxicity following exposure to CS, providing an objective measure for the suitability of the cellular model. To evaluate the impact of various treatments on cellular Trans Epithelial Electrical Resistance (TEER) values, we utilized the control group as a baseline. We computed the differences in TEER values between different treatment groups and the control group to depict the extent of influence on cellular membrane integrity.
5
Transendothelial Electrical Resistance Measurement for Cell Monolayers
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The TEER was monitored for 1 h at different time points (0, 10, 30, 45, 60 min) in SFM medium using the cultures of HBEC and hiPS-BMEC seeded in the 24-well insert. The TEER was measured using a Millicell® ERS-2 Volt-Ohm Meter (Millipore, Bedford, MA, USA) equipped with a STX01 chopstick electrode (Millipore, Bedford, MA, USA). The TEER value was calculated from the following equation (1) [36 (link)]: Where Rmonolayer is the resistance of the cell monolayer along with the filter membrane; Rblank is the resistance of the cell-free insert membrane in SFM medium, and A is the surface area of the membrane (0.33 cm2).
6
Evaluating Cellular Barrier Integrity via TEER
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Trans-endothelial electrical resistance (TEER), an electrical parameter to assess membrane integrity and suitability of in vitro cellular barriers31 (link), was used to assess cell layer integrity and toxicity of CS exposure. TEER was measured using a Millicell-ERS-2 (Millipore, Billerica, MA, US) volt-ohm-meter with a STX01 chopstick electrode (Millipore, Billerica, MA, US). Triplicate measurements were performed for each well. Three wells per experimental run were tested. Subsequently, the values were adjusted with blank control. The obtained value was multiplied by the effective membrane area in cm2 (1.12 cm2 for 12-well transwell inserts) to yield the final result in Ω.cm2.
7
Measuring Epithelial Barrier Function
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TEER development of differentiating pHBECs was monitored using a Millicell-ERS-2 (Millipore; Billerica, MA) volt-ohm-meter with a STX01 chopstick electrode (Millipore)24 (link). For this, 500 μl pre-warmed HBSS was added to the apical compartment and left to equilibrate for 10 minutes in the incubator. Subsequently, a triplicate measurement was performed for every well. For different treatment conditions, 6 to 12 individual wells were analysed per experiment and time point. The values measured were corrected for blank values and area. For this, the average resistance of a blank well (without cells) was subtracted from the measured value of every well. The obtained value was multiplied by the effective membrane area in cm2 (1.12 cm2 for 12-well transwell inserts) to yield the final result in Ω x cm2. After TEER measurement, the apical solution was removed to restore ALI culture conditions.
8
Measuring Transepithelial Electrical Resistance in 3D BBB Model
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The transepithelial electrical resistance (TEER) was measured for the fibrin gel without BBB cells and for 3D BBB model with open structures after 7 days culture. The TEER was measured in PBS at RT using a Millicell® ERS-2 Volt-Ohm Meter (Millipore, Bedford, MA, USA) equipped with a STX01 chopstick electrode (Millipore, Bedford, MA, USA). The TEER value was calculated from the following equation (1) [31 (link)]: where Ronly gel is the resistance of the fibrin gel without BBB cells; RBBB is the resistance of the fibrin gel seeded with BBB cells, and A is the average value of CD31+ surface area of three 3D BBB gels calculated by IMARIS software.
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