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bioRxiv · 10.1101/2023.03.26.534303

A versatile high-throughput assay based on 3D ring-shaped cardiac tissues generated from human induced pluripotent stem cell derived cardiomyocytes

Seguret, M.·Davidson, P.·Robben, S.·Jouve, C.·Pereira, C.·Cerveau, C.·Le Berre, M.·Rodrigues Ribeiro, R. S.·Hulot, J.-S.
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Abstract

We developed a 96-well plate assay which allows fast, reproducible and high-throughput generation of 3D cardiac rings around a deformable optically transparent hydrogel (PEG) pillar of known stiffness. Human induced pluripotent stem cell-derived cardiomyocytes, mixed with normal human adult dermal fibroblasts in an optimized 3:1 ratio, self-organized to form ring-shaped cardiac constructs. Immunostaining showed that the fibroblasts form a basal layer in contact with the glass, stabilizing the muscular fiber above. Tissues started contracting around the pillar at D1 and their fractional shortening increased until D7, reaching a plateau at 25{+/-}1%, that was maintained up to 14 days. The average stress, calculated from the compaction of the central pillar during contractions, was 1.4{+/-}0.4 mN/mm2. The cardiac constructs recapitulated expected inotropic responses to calcium and various drugs (isoproterenol, verapamil) as well as the arrhythmogenic effects of dofetilide. This versatile high-throughput assay allows multiple in situ mechanical and structural read-outs.

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Seguret, M., Davidson, P., Robben, S., Jouve, C., Pereira, C., Cerveau, C., Le Berre, M., Rodrigues Ribeiro, R. S., Hulot, J.-S.. 2023-03-29. A versatile high-throughput assay based on 3D ring-shaped cardiac tissues generated from human induced pluripotent stem cell derived cardiomyocytes. https://doi.org/10.1101/2023.03.26.534303

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