cat no | 02-1001-0000
myrCell Ventricular Cardiomyocytes
Human iPSC-derived ventricular cardiomyocytes
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Cryopreserved human iPSC-derived ventricular cardiomyocytes generated by directed differentiation that are ready for experiments in days
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Ideal for studying cardiovascular disease and cardiotoxicity in 2D or 3D engineered heart muscle
- Highly characterised to show functional synchronised beating by day 7 post-revival
Human iPSC-derived ventricular cardiomyocytes
Immunofluorescent staining of human iPSC-derived Ventricular Cardiomyocytes 10 days post-thawing showing expression of α-actinin (structural protein in the Z-discs, responsible for organised contractile sarcomere; green), and cardiac troponin I (TNNI3) (a cardiac-specific component of the troponin complex involved in regulation of myocardial contraction and a marker of differentiated ventricular cardiomyocytes; red) and nuclei stained with DAPI (blue). 60x magnification.
Ventricular Cardiomyocytes form confluent, electrically coupled monolayers by day 12
Brightfield images showing the progressive recovery and attachment of Ventricular Cardiomyocytes cultured on Matrigel-coated flasks. Cells form confluent, electrically coupled monolayers and develop spontaneous, synchronised beating following thawing. Images were acquired at days 1, 4, and 12 post-thaw (10x objective).
Ventricular Cardiomyocytes express key cardiac specific markers
Representative immunofluorescent staining of human iPSC-derived ventricular cardiomyocytes at 10 days post-thaw showing co-localisation of cardiac troponin T (cTnT) with α-actinin, and cTNT with mitochondria (mtCOX2); 60x magnification.
RNA sequencing confirms distinct ventricular and atrial phenotypes
RNA sequencing (RNA-seq) demonstrates robust chamber-specific differentiation of Ventricular and Atrial Cardiomyocytes (CM). Differential gene expression and hierarchical clustering reveal distinct ventricular and atrial transcriptomic signatures, confirming reproducible lineage specification and molecular identity across independent differentiation batches. The cells were harvested for RNA-seq at day 7 post-thaw.
Volcano plot showing differential gene expression. The x-axis represents log2 fold change (FC) and the y-axis shows -log10 (p value) for each gene. Genes with significant upregulation (blue) or downregulation (red) are labeled, with prominent cardiac-related genes (A).
Heatmap depicting the relative expression (row Z-score) of selected cardiac-related genes across samples; gene names are listed on the right. The heatmap reveals distinct clustering patterns between conditions. The colour scale ranges from -3 (red, indicating lower expression) to +3 (blue, indicating higher expression) (B).
Data were generated from three wild-type iPSC lines (W001, W003, W005) differentiated into both ventricular and atrial cardiomyocytes across 3–4 independent batches. For each line and batch, three technical replicates were pooled prior to sequencing. Ventricular Cardiomyocytes, cat. no. 02-1001-0000, and Atrial Cardiomyocytes, cat. no. 06-1001-0000, are generated from iPSC line W001. Please enquire for further information about iPSC lines W003 and W005.
Robust functional expression of key cardiac ion channels confirmed by automated patch-clamp analysis
Automated patch-clamp analysis confirms robust expression of key cardiac ion channels in Ventricular (red) and Atrial (blue) Cardiomyocytes (CM).
Representative INa current traces with mean ± SEM peak INa amplitude and I-V curve (A).
Representative ICa,L current traces with mean ± SEM peak ICa,L amplitude and I-V curve fitted with Boltzmann-Ohm curve (B).
Representative trace showing basal inward rectifier current (IK1) in the presence of 20 mM KCl (black) and BaCl2 (grey) and mean ± SEM IK1 at -100 mV (C).
Measurements were acquired from iPSC-derived cardiomyocytes cultured for 50 days post-thaw. Comparisons were made using unpaired Student’s t-test, Mann Whitney’s U-test vs. Ventricular. ****p<0.0001, *p<0.05; n/N = iPSC-CM/batches.
Mature electrophysiological phenotypes demonstrated by action potential recordings
Representative action potential (AP) recordings demonstrate mature ventricular and atrial phenotypes.
Representative membrane voltage (mV) traces showing atrial and ventricular triggered APs in Atrial (blue) and Ventricular (red) Cardiomyocytes (CM) (left).
AP duration at 90% (APD90) and 50% (APD50) repolarisation as mean ±SEM (centre and right).
Measurements were acquired from human iPSC-derived cardiomyocytes cultured for 7 days post-thaw. Comparisons were made using unpaired Student's t-test, Mann Whitney's U-test vs. Ventricular. ****p<0.0001; n = iPSC-CM from one batch.
Chamber-specific contractile properties confirmed by quantitation of contractile dynamics
Contraction analysis using CytoMotion label-free, image-based motion detection software (IonOptix), confirms chamber-specific contractile function of Ventricular (red) and Atrial (blue) Cardiomyocytes (CM).
Frequency (Hz), time to peak (sec), and time to 90% baseline (sec) showed as mean ± SEM (A).
Representative traces showing contraction of human iPSC-derived cardiomyocytes monolayers as mean ± SEM (B).
Measurements were acquired from iPSC-derived cardiomyocytes cultured for 50 days post-thaw. Comparisons were made using unpaired Student's t-test, Mann Whitney's U-test vs. Ventricular. ****p<0.001; n/N = measurement ROI/batches.
Mitochondrial stress test confirms metabolically mature bioenergetics
The Seahorse XF Mito Stress Test demonstrates robust mitochondrial respiration and oxidative metabolism in wild-type Ventricular Cardiomyocytes (WT). Cells were maintained in continuous culture and analysed at two maturation stages (Day 21 and Day 31). Seven days prior to the assay (Day 14 and Day 24), cells were seeded into XFe24 cell culture plates at 80,000 cells per well. On the day of the experiment, mitochondrial function was assessed using the Seahorse XF Mito Stress XF RPMI medium, pH 7.4 supplemented with 10 mM Glucose, 1 mM pyruvate, 200 µM L-AsC, 2% B27.
Representative trace of changes in oxygen consumption rate in Ventricular Cardiomyocytes when treated with compounds affecting the mitochondrial electron transport chain, showed as mean ± SEM (A).
Mitochondrial respiration, spare respiratory capacity, ATP production and efficiency derived from the representative trace displayed in (A), shown as mean ± SEM (B).
As cardiomyocytes mature from Day 21 to Day 31, they develop enhanced mitochondrial function and bioenergetic capacity, evidenced by increased mitochondrial respiration, higher ATP-linked respiration, and greater spare respiratory capacity.
Comparisons were made using unpaired Student’s t-test, *p<0.05, **p<0.01, n = 5.
Engineered heart muscle exhibits progressive maturation and a positive force-frequency relationship
Human iPSC-derived ventricular cardiomyocytes were combined with fibroblasts in a collagen-based hydrogel and cast into myrPlates to generate Engineered Heart Muscle (EHM). Cardiomyocytes were cultured for 7-10 days post thaw before tissue casting. Live video-optical tracking using automated pole detection and peak analysis in the myrImager platform enabled longitudinal assessment of tissue contractility throughout maturation.
Immunofluorescent imaging of the cross section shows a compact, organised tissue with aligned cardiomyocytes, structured sarcomeres and abundant mitochondria.
Engineered heart muscle exhibits progressive maturation and a positive force-frequency relationship
Human iPSC-derived ventricular cardiomyocytes were combined with fibroblasts in a collagen-based hydrogel and cast into myrPlates to generate Engineered Heart Muscle (EHM). Cardiomyocytes were cultured for 7-10 days post thaw before tissue casting. Live video-optical tracking using automated pole detection and peak analysis in the myrImager platform enabled longitudinal assessment of tissue contractility throughout maturation.
Video-optical measurements of 16 EHMs over 42 days demonstrated progressive increases in force of contraction (FOC), consistent with continued functional maturation (B). Resting length, quantified as the diastolic pole-to-pole distance in myrPlate cultures, provides a longitudinal readout of tissue compaction and mechanical state. Data are shown as mean ± SEM, n=16.
At study endpoint, EHMs were subjected to isometric organ-bath analysis to independently characterise and validate tissue contractile properties (C). The upper panel shows the preload-dependent Frank–Starling response during 1.5 Hz electrical stimulation at 1.8 mM extracellular Ca2+. Stepwise increases in preload assess the relationship between initial muscle length and force development, a fundamental property of functional myocardium. The lower panel shows the calcium-dependent inotropic response, measured during 1.5 Hz stimulation across increasing extracellular Ca2+ concentrations (0.2–4 mM), providing a measure of maximal calcium-dependent force-generating capacity. Data are shown as mean ± SEM, n=6–12.
Engineered heart muscle exhibits metabolic flexibility and oxidative maturation
Engineered Heart Muscle (EHM) was evaluated using Seahorse XF metabolic stress and fuel dependency assays (Agilent Technologies) to characterise mitochondrial respiration and substrate use. EHMs display robust oxidative metabolism and metabolic flexibility by using glucose, fatty acids, and alternative substrates for ATP production, reflecting progressive metabolic maturation toward an adult-like cardiac phenotype. EHMs were cultured for 42 days before transferring one EHM per well to a Seahorse XFe24 tissue culture plate. A mitochondrial stress test or fuel dependency test was performed to investigate mitochondrial function and metabolite use. Ventricular Cardiomyocytes were harvested from a 25-day continuous culture before EHM casting.
Representative trace of changes in oxygen consumption rate in EHM when treated with compounds affecting the mitochondrial electron transport chain, shown as mean ± SEM (left).
Representative trace of changes in oxygen consumption rate in EHM when treated with compounds inhibiting processing of glucose (2-deoxy-D-glucose), inhibiting uptake of fatty acids into mitochondria (etomoxir), or inhibiting the electron transport chain, with data shown as mean ± SEM (centre).
Metabolic fuel dependency derived from data in centre shows increased use of fatty acids compared to glucose. Data shown as mean ± SEM (right).
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