Abhishek P Dhand, Miranda A Juarros, Thomas G Martin, Manuela Garay-Sarmiento, Christina Alamana, Dakota R Hunt, Gabriel J Rodriguez-Rivera, Mackenzie C Obenreder, Cody O Crosby, Bianca Meurer-Zeman, Quentin McAfee, Henry Valle-Ayala, Hannah M Zlotnick, Declan N Goddard, Christopher C Ebmeier, Jason A Burdick, Leslie A Leinwand
Investigations into cardiac biology and drug discovery benefit from in vitro models that replicate human cardiac physiology. Current engineered heart tissue (EHT) models recapitulate aspects of this physiology but are limited by scalability, cost, and reproducibility. We report a one-step method to fabricate hydrogel molds using digital light processing (DLP) 3D printing to support EHT formation from human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). DLP enables rapid tuning of mold size, aspect ratio, and stiffness, while increasing scalability for high-throughput experiments without compromising EHT quality. DLP-fabricated EHTs show aligned extracellular matrix and display improvements in maturity versus 2D cultures, including a shift to fatty acid metabolism, increased myofilament density, and improved sarcomere organization. These EHTs also display expected responses to pathological stimuli (e.g., adrenergic agonism, increased stiffness), enabling disease modeling applications. Overall, 3D DLP-printed EHT molds provide versatile, scalable cardiac tissue platforms for both mechanistic studies and large-scale drug screening.