cat no | 02-5001-0010
myrCell SarcPaint Ventricular Cardiomyocytes TTN A-band mutation
Human iPSC-derived ventricular cardiomyocytes with a TTN mutation and live sarcomere reporter
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Cryopreserved human iPSC-derived cardiomyocytes with a TTN A-band frameshift mutation and ACTN2-Citrine reporter, generated by directed differentiation, and ready for experiments in days
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Reporter enables live, longitudinal visualisation of disease-specific sarcomere organisation and disarray
- Genetically matched disease model for quantitative phenotyping of TTN-associated dilated cardiomyopathy in 2D and 3D tissues
Human iPSC-derived ventricular cardiomyocytes with a TTN A-band mutation and live sarcomere reporter
Immunofluorescence staining of human iPSC-derived SarcPaint Ventricular Cardiomyocytes TTN A-band mutation 10 days post-thawing showing endogenous α-actinin-Citrine reporter (green), cTNT (magenta), α-actinin (red), and nuclei stained with DAPI (blue). Alpha-actinin marks the sarcomeric structure, while cTNT confirms cardiac identity. The TTN A-band mutant cardiomyocytes show a disturbed sarcomeric organisation, consistent with the disease-associated phenotype. 60x magnification.
SarcPaint Ventricular Cardiomyocytes TTN A-band mutation express key cardiac specific markers
Immunofluorescent staining of human iPSC-derived Ventricular Cardiomyocytes carrying a TTN A-band frameshift mutation and genetically encoded live sarcomere reporter at 10 days post-thawing. SarcPaint-tagged α-actinin is observed in all cells, with co-localisation of cardiac HCN4, cardiac troponin T (cTnT), or mitohondria (mtCOX2); 40x magnification.
Progressive disruption of sarcomere organisation and contractility in TTN A-band mutant ventricular cardiomyocytes
The SarcPaint reporter enables comparison of functional sarcomere organisation across parental wild-type (WT) ventricular cardiomyocytes and disease models carrying a heterozygous (HET) or homozygous (HOM) TTN A-band frameshift mutation (c.70692_70693insAT). Note: the WT and patient-relevant HET models are available as products, whereas the non-naturally occurring HOM model was included specifically to demonstrate a more severe phenotype (enquire for further information).
A) WT, HET, and HOM SarcPaint cardiomyocytes were seeded into a 384-well plate and periodically recorded over 11 days.
B) Spontaneous cardiomyocyte contractions recorded at day 7.
Contractions are clearly detectable in WT cells, visibly reduced in the HET model, and barely detectable in the HOM model. Overall, the disease models exhibit progressively disrupted sarcomere organisation and contractility, demonstrating the utility of SarcPaint for visual assessment and downstream quantitative phenotyping. Scale: 60x magnification.
Functional assessment of WT and TTN A-band mutant engineered heart muscle
Wild-type (WT) or TTN A-band mutant (TTN mutant) Ventricular Cardiomyocytes expressing the SarcPaint reporter were combined with fibroblasts in a collagen-based hydrogel to generate engineered heart muscle (EHM). The cells were maintained for 7-10 days post-thaw before being cast into myrPlates. Live video-optical tracking, using automated pole detection and peak analysis in the myrImager, enabled longitudinal assessment of tissue contractility throughout maturation.
Longitudinal video-optical contractility measurements of 16 EHM tissues over 35 days of culture (left). The data demonstrate a progressive improvement in the force of contraction (FOC) in WT EHMs, whereas TTN mutant EHMs exhibit a compromised FOC, indicative of impaired tissue function.
TTN mutant EHMs exhibited reduced resting tension compared with WT tissues, indicating altered passive mechanical properties associated with the TTN mutation (right). Data shown as mean ± SEM, n=16.
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