Method Profile
A single human iPSC line supplies three cardiac cell types for matched endothelial-containing and endothelial-free drug tests under perfusion.
Investigate how endothelial inclusion changes acute contractile and structural drug responses.
Human iPSC line FiPSCtrl1-mR5F-6-derived cardiomyocytes, cardiac fibroblasts and endothelial cells; 90:10 CM:CF on one PET-membrane face and EC on the other.
Four 0.32 cm² chambers; shared-reservoir perfusion 5 μL/min; seven-day culture baseline, one-hour isoproterenol and 24-hour doxorubicin exposure.
Culture controls
PET versus polycarbonate attachment, 90:10 versus 50:50 CM:CF, gelatin versus fibronectin for EC; EGM2 medium for tri-culture. Later fibroblast passage caused cardiac-marker drift.
Drug comparisons
Isoproterenol 1 μM for one hour and doxorubicin 1 μM for 24 hours introduced through lower channel. Separate 2D cardiomyocyte doxorubicin 0.01–1,000 μM viability curve, IC50 = 8.7 μM.
Contractile changes are normalized to baseline and compared between matched bi- and tri-cultures. The separate 2D viability IC50, chip functional responses and endothelial tracer measurements assess different features.
Isogenic endothelial-containing versus endothelial-free cardiac culture under matched acute drug challenge.
One iPSC line; three independent experiments; contractility n = 5 CM/CF and n = 6 tri-culture measurements. Sarcomere comparisons use their separate reported sampling.
CM/CF versus CM/CF/EC; baseline/vehicle; isoproterenol versus doxorubicin.
Isoproterenol increased beating similarly in both configurations, with other kinetic differences nonsignificant. Doxorubicin reduced endothelial-free beating by 72% and disrupted sarcomeres; tri-culture preserved cardiac responses despite endothelial junction disruption.
A heart chip combined cardiomyocytes, cardiac fibroblasts and endothelium from one human iPSC line. Endothelial-containing cultures preserved beating and sarcomere organization during 1 μM doxorubicin exposure, while isoproterenol increased beating similarly with or without endothelium.
