Characterization & Applications of Human Cardiomyocytes

ReproCardio 2 Human iPSC-derived cardiomyocytes have been extensively characterized for their functional responsiveness to the known cardiotoxic compounds. In all our tests, including assays of cardiotoxic compounds not correctly identified by the hERG assay, ReproCardio has generated results in complete agreement with clinical findings.

In this post, we’ll highlight the characterisation of ReproCardio 2 cardiomyocytes and illustrate the applications they are used for.

Characterization of ReproCardio 2 Cardiomyocytes

ReproCardio 2 cardiomyocytes are provided frozen, but even after thawing they retain structural integrity and function thanks to the specially developed freezing technology.

Expression of various cardiomyocyte-specific marker genes and cardiac ion channels

The expression of various cardiomyocyte-specific marker genes and cardiac ion channels have been confirmed. Moreover, ReproCELL’s specially-developed freezing technology makes it possible to keep a beautiful cytoskeleton after thawing of the frozen cells, which is not trivial for cardiomyocytes. (Blue:DAPI)

Expression of heart cell markers and structural integrity after freeze-thaw.

Expression of heart cell markers and structural integrity after freeze-thaw.

 

Electrophysiological characterization with patch clamp

ReproCardio2 can be used for manual patch clamp experiments when cultured as single cells. In vivo-like activities and properties including spontaneous action potential, Na/Ca/K channel currents are reliably observed. ReproCardio Coat for Patch Clamp is highly recommended for patch clamp assays.

 

*ReproCardio Coat for Patch Clamp is highly recommended for patch clamp assays. ReproCardio2 can be used for manual patch clamp experiments when cultured as single cells. In vivo-like activities and properties including spontaneous action potential, Na/Ca/K channel currents are reliably observed.

Electrophysiological characterization with patch clamp.

Applications for ReproCardio 2 Cardiomyocytes

MEA recordings from beating thin layer formed by ReproCardio 2

When plated on a flat surface coated with ReproCELL’s coating material provided in the kit, ReproCardio 2 cardiomyocytes form a thin layer and start beating. Extracellular electrical recordings can be made by using MEA, and clear sodium and potassium peaks can be observed.

Technical tips icon by tebu-bioWe recommend that the CO2 concentration (5%) is fixed during measurement with MEA for acquiring stable data.

The following film represents MEA recording from a thin layer constituted of ReproCardio 2 cardiomyocytes

 

 Extracellular electrical activity. Red vertical lines indicates sodium and potassium signals.

Extracellular electrical activity. Red vertical lines indicates sodium and potassium signals.

 

Live imaging of calcium concentration change using ReproCardio 2 thin layer

Calcium wave propagation over the cardiomyocyte thin layer can be observed by using conventional calcium-sensitive dyes. The accessibility of the cells within a thin layer for drugs makes this suitable for testing the effects of compounds to calcium activity of cardiomyocytes.

The film represents calcium imaging of a thin layer constituted of ReproCardio 2 cardiomyocytes using a calcium-sensitive dye (fluo-8).

 

Quantification of fluorescent intensity which represents calcium dynamics in the cells

Quantification of fluorescent intensity which represents calcium dynamics in the cells

 

Forming multicellular clusters from ReproCardio2

Clusters can be formed easily by using the 96 well plate included in the kit.

 

A beating cardiomyocyte cluster on MEA

 

ReproCardio 2 cardiomyocyte clusters show robust beating. Extracellular electrophysiological recordings can be made by placing the clusters on electrodes like those of MEA system.

 

in vitro ECG-like waveform resembling in vivo heart activity

ReproCardio 2 cardiomyocyte clusters, when extracellular recording is performed, can generate ECG-like waveform. The effects of various compounds can be functionally evaluated using this in vivo-like system.

The assay medium for extracellular recordings and live imaging is available separately.

So, what do you think of these ReproCardio 2 cardiomyocytes? Share your comments or questions below!

 

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  1. […] In a previous post, I highlighted the characterisation of ReproCardio 2 cardiomyocytes and illustrated the applications they are used for. […]

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