Cardiotoxicity assessment using 3D vascularized cardiac tissue consisting of human iPSC-derived cardiomyocytes and fibroblasts

Figures

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  Figure 1. Immunostaining of hiPSC-derived 3D cardiac tissues Cardiac cell type (A) CD31 (red), (B) vimentin (green) and cardiac troponin T (red), cardiac-specific structural proteins: (C) sarcomeric α-actinin (red) and connexin43 (green), (D) MLC2v (green) and MLC2a (red), and extracellular matrix proteins: (E) fibronectin (red) and cardiac troponin T (green), (F) laminin (green) and cardiac troponin T (red), (G) collagen type I (green) and cardiac troponin T (red). Nuclei were stained with DAPI. The fluorescent images were acquired at 10 (A) or 60 (B–G) magnifications.

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  Figure 2. Baseline values of motion parameters acquired with motion vector analysis All individual values before treatment with test compounds or vehicle were plotted for each parameter. Bar and error bars indicate the mean ± SD (n = 72).

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  Figure 3. Effects of DMSO on contraction and relaxation waveform (A) Examples of a typical motion waveform are represented before (baseline) and after four times treatment with 0.03% DMSO in hiPSC-derived 3D cardiac tissues. (B) Effects of 0.03% DMSO on motion parameters acquired from motion vector analysis. The values are represented as mean ± SD (n = 6) of the percentage of baseline after treatment with DMSO compared to medium-treated values.

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  Figure 4. Effects of test compounds on contraction and relaxation waveform Representative examples of motion waveforms are represented before (baseline) and after treatment with isoproterenol (1–30 nM or 30–1,000 nM), dobutamine (3–100 μM), milrinone (3–100 μM), ouabain (3–30 nM), pimobendan (0.1–3 μM), glibenclamide (1–30 μM), verapamil (3–100 nM), flecainide (30–1,000 nM), E-4031 (3–30 nM), and chromanol 293B (1–30 μM) in hiPSC-derived 3D cardiac tissues.

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  Figure 5. Effects on beating rate of test compounds affecting cardiac contractility and repolarization Data are expressed as means ± SD of 6 preparations. ∗p < 0.05, ∗∗p < 0.01 compared with baseline (one-way ANOVA followed by Dunnett post hoc test). The gray shaded area shows mean and 2 SD of percentage of baseline after treatment with vehicle.

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  Figure 6. Effects on contractile and relaxation parameters of test compounds Maximum contraction speed, black, closed circle; maximum relaxation speed, blue, closed triangle; average deformation distance, red, closed square. Data are expressed as means ± SD of 6 preparations. ∗p < 0.05, ∗∗p < 0.01 compared with baseline (one-way ANOVA followed by Dunnett post hoc test).

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  Figure 7. Effects on repolarization parameters of test compounds Relaxation time, closed circle; contraction-relaxation duration, closed square. Data are expressed as means ± SD of 6 preparations. ∗p < 0.05, ∗∗p < 0.01 compared with baseline (one-way ANOVA followed by Dunnett post hoc test). The gray shaded area shows mean and 2 SD of percentage of baseline after treatment with vehicle.