Extended Data Fig. 2: Functional comparison of hiPSC-CMs and human cardiac organoids.
From: Electro-metabolic coupling in multi-chambered vascularized human cardiac organoids
a, Principal component analysis (PCA) of 513 genes differentially expressed between hiPS-derived cardiomyocytes and vascularized cardiac organoids (methods). Cardiac organoids cluster with adult, but not fetal cardiomyocytes. b, Principal component analysis (PCA) of 513 genes differentially expressed between hiPS-derived cardiomyocytes and vascularized cardiac organoids (methods) separated into PC components. PC1 gene set is enriched for angiogenesis and cell adhesion, clustering the non-beating AC16 cell line, the negative control, with the mature heart tissue. This clearly suggests that PC1 is less relevant for comparative analysis. c, Visual contraction analysis of UN-1 vascularized cardiac organoids treated with DMSO (Control), 10 µM amiodarone, or 100 µM epinephrine (Supplementary. Video 3), normalized to the highest and lowest signal recorded through the entire measurement duration (30 seconds; methods). Analysis shows that untreated organoids acquire a homogenous synchronized spontaneous beating of 66 ± 5 beats per minute. Stimulation with 100 µM epinephrine increases the contraction rate to 88 ± 7 bpm and relative contraction by 18% (n = 5, p < 0.001), while stimulation with 10 µM amiodarone decreased the rate to 52 ± 4 bpm and contraction by 28% (n = 5, p < 0.001), resulting in a physiological-like response to the drugs. Mean of 5 biological replicates; error bars, s.e.m. Significance was determined using a one-way ANOVA with Dunnett correction. Graphs were taken from Fig. 3d. d, Visual contraction analysis of UN-1 hiPSC-derived cardiomyocytes (hiPSC-CMs) treated with DMSO (Control), 10 µM amiodarone, or 100 µM epinephrine. Analysis shows that untreated hiPSC-CMs acquire a homogenous unsynchronized beating of 97 ± 4 beats per minute. Stimulation with 100 µM epinephrine increases the contraction rate to 106 ± 4 bpm and relative contraction by 18% (n = 3, p < 0.01), while stimulation with 10 µM amiodarone decreased the rate to 92 ± 3 bpm and contraction by 28% (n = 3, p < 0.01). Mean of 3 biological replicates; error bars, s.e.m. Significance was determined using a one-way ANOVA with Dunnett correction. e, Seahorse MitoStress assay nested analysis of UN-1 cardiac organoids, UN-1 hiPSC-derived cardiomyocytes (hiPSC-CMs), and cardiac endothelial cells (CECs). CECs show basal respiration equal to 4.5% of the basal respiration of the cardiac organoids and less than 2% of maximal respiration, indicating that changes in respiration are attributed to changes in the cardiomyocytes (n = 9, p < 0.001). Significance was determined using one-way ANOVA and Dunnett multiple comparison correction. Lines represent independent experiments, Error bars mark standard error of mean among n = 3 biological repeats.
