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Understanding the structural and functional development of human-inducedpluripotent stem-cell-derived cardiomyocytes is essential to engineeringcardiac tissue that enables pharmaceutical testing, modeling diseases, anddesigning therapies. Here we use a method not commonly applied to biologicalmaterials, small angle X-ray scattering, to characterize the structuraldevelopment of human-induced pluripotent stem-cell-derived cardiomyocyteswithin 3D engineered tissues during their preliminary stages of maturation. AnX-ray scattering experimental method enables the reliable characterization ofthe cardiomyocyte myofilament spacing with maturation time. The myofilamentlattice spacing monotonically decreases as the tissue matures from its initialpost-seeding state over the span of ten days. Visualization of the spacing at agrid of positions in the tissue provides an approach to characterizing thematuration and organization of cardiomyocyte myofilaments and has the potentialto help elucidate mechanisms of pathophysiology, and disease progression,thereby stimulating new biological hypotheses in stem cell engineering.

Structural maturation of myofilaments in engineered 3D cardiac microtissues characterized using small angle X-ray scattering