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The structural and functional maturation of human induced pluripotent stemcell-derived cardiomyocytes (hiPSC-CMs) is essential for application topharmaceutical testing, disease modeling, and ultimately therapeutic use.Multicellular 3D-tissue platforms have improved functional maturation ofhiPSC-CMs, but probing cardiac contractile properties remains challenging in a3D environment, especially at depth and in live tissues. Using small angleX-ray scattering (SAXS) images, we show that hiPSC-CMs, matured and examined ina 3D environment, exhibit periodic spatial arrangement of the myofilamentlattice, which has not been previously detected in hiPSC-CMs. Contractile forceis found to correlate with both scattering intensity (R2=0.44) and latticespacing (R2=0.46). Scattering intensity also correlates with lattice spacing(R2=0.81), suggestive of lower noise in our structural measurement relative tothe functional measurement. Notably, we observe decreased myofilament orderingin tissues with a myofilament mutation known to lead to hypertrophiccardiomyopathy (HCM). Our results highlight the progress of human cardiactissue engineering and enable unprecedented study of structural maturation inhiPSC-CMs.

Probing Subcellular Nanostructure of Engineered Human Cardiomyocytes in 3D Tissue