Short-circuit current measurements were made essentially as described in prior studies [8 (link), 9 (link)]. In brief, measurements were made using a DVC-1000 voltage clamp (World Precision Instruments Inc., Sarasota, FL), cells were cultured on Snapwell clear permeable supports (Corning), and experiments were performed using HCO3−-buffered solutions at 37 °C. For studies using FRT cells the basolateral membrane was permeabilized with 250 μg/ml amphotericin B and experiments were done with a chloride gradient. For 16HBE14o- gene-edited cells experiments were done with a chloride gradient using Hepes-buffered solutions.
Snapwell clear permeable supports
Manufactured by Corning
Snapwell clear permeable supports are a type of lab equipment designed for cell culture applications. They provide a clear, permeable surface that allows for the growth and study of cells in a controlled environment. The core function of these supports is to facilitate the culture and analysis of cells without obstructing visibility or hindering their growth.
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Lab products found in correlation
2 protocols using snapwell clear permeable supports
1
Short-circuit Current Measurement Protocol
2
Measurement of Short-Circuit Current in Airway Epithelial Cells
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Short-circuit current was measured on cells cultured on Snapwell clear permeable supports (Corning) as described10 (link),11 (link). Hemichambers were connected to a DVC-1000 voltage clamp (World Precision Instruments Inc., Sarasota, FL) via Ag/AgCl electrodes and 3 M KCl agar bridges for recording of the short-circuit current. For human airway epithelial cells, symmetrical HCO3−-buffered solutions (in mM: 120 NaCl, 5 KCl, 1 MgCl2, 1 CaCl2, 5 Hepes, 25 NaHCO3, 10 glucose, pH 7.4) for basolateral and apical side were used. For FRT cells, the basolateral membrane was permeabilized with 250 μg/ml amphotericin B, and experiments were done with a chloride gradient using a HCO3−-buffered solutions (Basolateral (mM): 120 NaCl, 5 KCl, 1 MgCl2, 1 CaCl2, 5 Hepes, 25 NaHCO3, 10 glucose, pH 7.4; apical (mM): 60 NaCl, 60 mM sodium gluconate, 5 KCl, 1 MgCl2, 1 CaCl2, 5 Hepes, 25 NaHCO3, 10 glucose, pH 7.4). All cells were equilibrated with 95% O2, 5% CO2 and maintained at 37 °C during experiments.
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