Kowalczewski, A.; Wang, C.; Wang, X.; Yang, H.; Qin, Z.; Ma, Z.

Understanding how cardiomyocyte structure governs contractile function is fundamental to cardiac biology and disease modeling, yet current approaches rely on time resolved imaging and computationally intensive analysis. Here, we present a generative artificial intelligence (AI) framework that directly predicts contractile behavior of human induced pluripotent stem cell-derived cardiomyocytes from single static images. Our approach integrates a U-Net-based generator with a patch-based generative adversarial network (GAN) discriminator to translate morphological and sarcomere structural features into pixel-resolved contraction heatmaps. This U-Net-GAN model achieved high predictive accuracy, with structural similarity index (SSIM) values up to 0.84 using combined morphological and structural inputs. To further enhance performance and generalizability, we incorporated synthetic cell function pairs generated via a generative AI StyleGAN2 framework, improving prediction accuracy and perceptual similarity. Importantly, region-specific and whole-cell analyses revealed that AI predictions capture biologically meaningful structure function relationships, with sarcomere organization strongly associated with both contractile output and prediction fidelity. Reconstruction error emerged as an interpretable metric reflecting localized inefficiencies in sarcomere-to-contraction coupling. Together, this framework establishes a scalable and interpretable strategy for inferring cardiomyocyte function from static morphology, eliminating the need for time-lapse imaging. More broadly, this work positions generative AI as a powerful tool for bridging cellular structure and function, enabling high-throughput functional phenotyping and advancing in vitro cardiac modeling.