Characterization of structural integrity, identity and function of ReproCardio 2

ReproCardio 2 human iPS cell-derived cardiomyocytes have been extensively characterized for their quality.

• Identity as the heart cells
• Use ReproCardio 2 as monolayers
• Use ReproCardio 2 as clumps
• Compound toxicity assays

 

Identity as the heart cells

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

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

MLC-2A Myosin light chain specific for the atrium of the mammalian heart.	MLC-2V Myosin light chain specific for the ventricle of the mammalian heart.	alpha-MHC Myosin heavy chain which is mainly expressed in the atrium of the heart.
beta-MHC Myosin heavy chain beta which is mainly expressed in the ventricle of the heart.	cTnT Cardiac troponin T, a component of troponin complex necessary for contraction	Cx43 Connexin 43, the main cardiac connexin found mainly in ventricular heart cells.

Use ReproCardio 2 as monolayers

Beating monolayer formed by ReproCardio 2 cardiomyocytes

When plated on a flat surface coated with ReproCELL’s coating material provided in the kit, ReproCardio 2 cardiomyocytes form a monolayer and start beating.

 

Live imaging of calcium concentration change using ReproCardio 2 cardiomyocyte monolayer

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

Calcium imaging of a monolayer constituted of ReproCardio 2 cardiomyocytes using a ratiometric calcium-sensitive dye (CC2DMPE -DisBac23, Invitorgen).

 

Use ReproCardio 2 as clumps

[Movie] Forming multicellular clusters from ReproCardio2

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

 

[Movie] A beating cardiomyocyte cluster on MEA

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

 

[Movie] in vitro ECG-like waveform resembling in vivo heart activity

ReproCardio 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.

 

Accurate replication of the in vivo response of the heart to cardiotoxic compounds

ReproCardio 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. For the detailed information about ReproCardio’s compound response, please refer to our web page for QTempo, an assay service for cardiotoxicity testing using ReproCardio cardiomyocytes.