hERG Current Recordings in BEL-7404 Cells

BEL-7404 cell lines overexpressing human ether-a-go-go-related gene (hERG) were used for the experiment (69 (link)). The cultured cells were placed on 35-mm diameter Petri dishes in extracellular solution and incubated at 37°C for 30 min to adhere. A whole-cell patch-clamp method was performed to record hERG currents at room temperature (22°C) using WPC-100 resistive feedback amplifiers (ESF electronic). When filled with internal pipette solution, the resistance of the patch pipettes (GC150F-10; Harvard Apparatus) averaged 2–3 MΩ. Patch pipettes were pulled from borosilicate glass by a P-97 horizontal puller (Sutter Instrument Co.). For drug application, PIX or cisapride (positive control) was applied using a piezoelectric-driven micromanipulator (P-287.70, Physik Instrumente) at a flow rate of 100–200 μL/min, as described previously (70 (link)).

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She P., Li Z., Li Y., Liu S., Li L., Yang Y., Zhou L, & Wu Y. (2022). Pixantrone Sensitizes Gram-Negative Pathogens to Rifampin. Microbiology Spectrum, 10(6), e02114-22.

Publication 2022

GC150F-10

Cell Cell lines Cisapride Cultured cells Dishes Drug Ether

Corresponding Organization : The First Hospital of Changsha

Other organizations : Third Xiangya Hospital, Central South University

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Variable analysis

independent variables

PIX or cisapride (positive control) applied using a piezoelectric-driven micromanipulator at a flow rate of 100–200 μL/min

dependent variables

HERG currents recorded using the whole-cell patch-clamp method

control variables

BEL-7404 cell lines overexpressing human ether-a-go-go-related gene (hERG)
Cells placed on 35-mm diameter Petri dishes in extracellular solution and incubated at 37°C for 30 min to adhere
Whole-cell patch-clamp method performed at room temperature (22°C) using WPC-100 resistive feedback amplifiers
Patch pipettes (GC150F-10; Harvard Apparatus) with resistance of 2–3 MΩ, pulled from borosilicate glass by a P-97 horizontal puller (Sutter Instrument Co.)

positive control

Cisapride

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