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 safety, 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
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.
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
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.
Technique
- Manual Patch Clamp (current clamp configuration)
- Non-cumulative concentrations
Study model
- Human cardiomyocytes
- 3 or 6 treated cells
Calcium current recordings
Sodium current recordings
Potassium current recordings
