Probing the subcellular nanostructure of engineered human cardiomyocytes in 3D tissue

Josh Javor; Jourdan K Ewoldt; Paige E Cloonan; Anant Chopra; Rebeccah J Luu; Guillaume Freychet; Mikhail Zhernenkov; Karl Ludwig; Jonathan G Seidman; Christine E Seidman; Christopher S Chen; David J Bishop

doi:10.1038/s41378-020-00234-x

Probing the subcellular nanostructure of engineered human cardiomyocytes in 3D tissue

Microsyst Nanoeng. 2021 Jan 27:7:10. doi: 10.1038/s41378-020-00234-x. eCollection 2021.

Authors

Josh Javor¹, Jourdan K Ewoldt², Paige E Cloonan², Anant Chopra², Rebeccah J Luu², Guillaume Freychet³, Mikhail Zhernenkov³, Karl Ludwig^{4

5}, Jonathan G Seidman⁶, Christine E Seidman⁶, Christopher S Chen^{1

2}, David J Bishop^{1

2

4

5

7}

Affiliations

¹ Department of Mechanical Engineering, Boston University, Boston, MA 02215 USA.
² Department of Biomedical Engineering, Boston University, Boston, MA 02215 USA.
³ SMI Beamline 12-ID, Brookhaven National Laboratory, Upton, NY 11973 USA.
⁴ Department of Physics, Boston University, Boston, MA 02215 USA.
⁵ Division of Materials Science, Boston University, Boston, Massachusetts 02215 USA.
⁶ Department of Genetics, Harvard Medical School, Boston, MA 02215 USA.
⁷ Department of Electrical Engineering, Boston University, Boston, MA 02215 USA.

Abstract

The structural and functional maturation of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is essential for pharmaceutical testing, disease modeling, and ultimately therapeutic use. Multicellular 3D-tissue platforms have improved the functional maturation of hiPSC-CMs, but probing cardiac contractile properties in a 3D environment remains challenging, especially at depth and in live tissues. Using small-angle X-ray scattering (SAXS) imaging, we show that hiPSC-CMs matured and examined in a 3D environment exhibit a periodic spatial arrangement of the myofilament lattice, which has not been previously detected in hiPSC-CMs. The contractile force is found to correlate with both the scattering intensity (R ² = 0.44) and lattice spacing (R ² = 0.46). The scattering intensity also correlates with lattice spacing (R ² = 0.81), suggestive of lower noise in our structural measurement than in the functional measurement. Notably, we observed decreased myofilament ordering in tissues with a myofilament mutation known to lead to hypertrophic cardiomyopathy (HCM). Our results highlight the progress of human cardiac tissue engineering and enable unprecedented study of structural maturation in hiPSC-CMs.

Keywords: Nanophotonics and plasmonics; Nanostructures; Organic-inorganic nanostructures.