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Heart disease is the leading cause of death in the US, and human cell models are needed to study how structure and function are related in heart health and disease. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are one promising cell model, but their immaturity (eg, lower sarcomere alignment and contractility) relative to adult human CMs limits their physiological relevance. While controlling cellular shape with protein micropatterning has enhanced maturity of single-cell hiPSC-CMs, they lack a key driver in cell maturity: cell-cell contacts. Here, we used a novel dual-protein patterning process to geometrically control single single-cell cardiomyocytes while maintaining the influence of cell-cell interactions. We first photopatterned two proteins on a glass coverslip using a two-step process with photomolecular adsorption of proteins. A laminin rectangle served to mimic the cell-ECM …