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Field Potential Changes in E-4031-Treated iCell Cardiomyocytes2 Using the CFPS-32 System

Date 2024.12

Author

Cellames Cell Analysis Team

Appendix

Field Potential Changes in E-4031-Treated iCell Cardiomyocytes2 Using the CFPS-32 System.pdf

Abstract

The purpose of this experiment is to analyze field potential changes in iCell cardiomyocytes2 treated with E-4031, a selective hERG potassium channel blocker.

Keyword

CFPS-32, iCell, Cardiomyocytes, E-4031, hERG, hERG potassium channel blocker, field potential, LPF

Main Text

Experiment Overview

Cell Name	iCell Cardiomyocytes²	Seeding quantity	35,000 cells
System	CFPS-32	Cell chip	CFPS-ITO-16W2E
Coating	Fibronectin 50mg/mL	Treatment	E-4031 (0.003, 0.01, 0.03, 0.1 mM)

Workflow

DIV0-7

DIV8 – Drug Treatment and recording protocol

- Plate iCell Cardiomyocytes² on Fibronectin-coated plates.

- Replace half of maintenance media every 48 hours until DIV7

- 20 μl/well of the 10X compound solutions were rapidly transferred to the MEA plate, followed by gentle mixing by pipetting 3~5 times. After 30 minutes, measurements were recorded for 5 minutes.

Results

Drug and concentration selection

Drug	Concentration (mM)
Drug	Dose1	Dose2	Dose3	Dose4
E-4031	0.003	0.01	0.03	0.1

Table1. The treatment concentrations of the E-4031

Field potential duration (FPD) is indicative of the activity of Ca²⁺ channels and hERG channels. A decrease in FPD indicates inhibition of the Ca²⁺ channels, whereas an increase in FPD suggests the inhibition of hERG channels. In this study, the selective hERG potassium channel blocker E-4031 was used to observe changes in the field potential waveform, and the alterations in parameters such as BPM, spike amplitude, and FPD were analyzed.
The treatment concentrations were selected (Table1.) based on those recommended by CiPA and the literature (J.R. Green et al., 2011, J Pharmacol Toxicol Methods).

Analysis of Cardiotoxicity following Drug treatment in iCell Cardiomyocytes2

T-test, GraphPad Prism (ver.10)
Statistical significance was considered at p < 0.05 (#), p < 0.01 (##), p < 0.001 (###), p < 0.0001 (####) and all p-values are reported accordingly.

Analysis of Field Potential Changes in iCell Cardiomyocytes2 with E-4031

A total of 35,000 iCell cardiomyocytes2 per well were cultured on CFPS-ITO-16W2E cell chip until DIV8, and field potentials before and after E-4031 treatment were compared and analyzed.
(A) The field potential waveform showed an increase in the beat period with higher doses of E-4031. At the highest dose (Dose 4), quiescence, indicating cardiac arrest, was observed.
(B) BPM decreased in a dose-dependent manner following E-4031 treatment. Compared to the control group, BPM was reduced by approximately 13.4% at Dose 1, 26.9% at Dose 2, and 59.8% at Dose 3.
(C) Spike amplitude decreased after E-4031 treatment, with a reduction of approximately 26.6% at Dose 3 compared to the control group.
(D) FPDcF was prolonged at higher doses of E-4031. EADs were observed in all tested wells (3/3) at Dose 4. Compared to the control group, FPDcF was prolonged by 47.1% at Dose 1, 95.7% at Dose 2, and 213% at Dose 3.

Summary

E-4031 treatment of iCell Cardiomyocytes2 cultured on CFPS-ITO-16W2E cell chips using the CFPS-32 system resulted in a dose-dependent increase in field potential duration, with reductions in BPM and spike amplitude, and prolonged FPDcF, while quiescence and EADs were observed at higher doses.

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Field Potential Changes in E-4031-Treated iCell Cardiomyocytes2 Using the CFPS-32 System