Human cardiomyocytes derived from iPS cells for preclinical drug studies

Healthy human heart cells are very rare and only available in very small quantities for obvious reasons of accessibility. Human cardiomyocytes derived from induced pluripotent stem cells (hiPSC-CM) allow to access human ventricular-like myocytes more easily. Due to their origin, these cells present the same functional and electrophysiological characteristics as human cardiomyocytes and offer a good alternative to ex vivo and in vivo models for drug development.

Human cardiomyocytes & cardiac safety studies

Human cardiomyocytes derived from iPS cells provide a good solution to study the repercussion of ion channels effects on cardiac electrical activity during preclinical studies. This human in vitro model can be used to evaluate the cardiac safety of newly developed drugs and is currently validated by the CiPA initiative for cardiac preclinical safety assessment.

What is shown?

To measure the effects of newly developed drugs on cardiac pharmacology, we study their action potential, i.e. we measure ion movements through the different transmembrane ionic channels. The modification of the shape of the action potential gives information about ion channel effects.

MEA impedance

Impedance measurements

Microelectrode array

MicroElectrode Array (MEA) recordings are a non-invasive way to measure integrated ion channel activity. The MEA system provides ECG-like results and detects modifications in beat rate, pro-arrhythmic events and other adverse effects.

Technique

xCELLigence RTCA Cardio ECR platform

Study model

Human IPSc derived cardiomyocytes
48 wells plate format
Simultaneous recording of MEA and impedance signals on the entire plate
Disease Models (Option)
- Brugada Syndrome
- Dilated Cardiomyopathy
- Hypertrophic Cardiomyopathy
- Catecholaminergic Polymorphic Ventricular Tachycardia

Protocol

1 negative control (DMSO 0.1%)
2 or 3 positive controls (1 concentration)
Compounds: several options from 1 compound (6 concentrations, n=6) up to 9 compounds (1 concentration, n=4)
Short-term and long-term exposure (up to 48 to 72h)

Measured parameters

Cell viability: Cell Index
Electrophysiology (MEA)
- Firing rate
- FPD
- FPDc (Fredericia)
- Spike amplitude
Contraction (Impedance)
- Amplitude of contraction
- Beat rate
- Beating period
- Individual beat duration (IBD10, 50, 90)
- Proarrhythmic events (BRI)

Reference compounds

Nifedipine
E-4031
Pentamidine

Results

Stability of recorded parameters in control conditions and reproductibility between plates

Typical effects of DMSO 0.1% on Beat rate.

Nifedipine (Low TdP risk)

Typical effects of Nifedipine on the amplitude of contraction.

Pentamidine (long-term exposure)

Typical effects of Pentamidine on impedance signal and FPDc.

Action potential recordings

Technique

Manual Patch Clamp (current clamp configuration)
Non-cumulative concentrations

Study model

Pluricyte® from Ncardia
3 or 6 treated cells

Measured parameters

Resting Potential (mV)
Amplitude (mV)
Action Potential Duration at 20% and 90% repolarisation : ADP₂₀ - ADP₉₀(ms)

Pluricyte® cardiomyocytes

Typical effects of nifedipine and bepridil on human cardiomyocytes derived from iPS (action potential recordings) on Pluricyte® cardiomyocytes.

Main cardiac ion channel recordings

The voltage-clamp technique allows to record the three specific currents (Ik, INa, ICa) that are involved in the action potential on individual human cardiomyocytes.