Self-assembly of hybrid 3D cultures by integrating living and synthetic cells

Self-assembly is a fundamental property of living matter that drives the three-dimensional (3D) organization of cell collectives such as tissues and organs. Here, the co-assembly of synthetic and natural cells is leveraged to create hybrid living 3D cancer cultures. We demonstrate that synthetic cells based on droplet-supported lipid bilayers can establish artificial tumor immune microenvironments (ART-TIMEs), mimicking immunogenic signals within tumoroids formed by the cell line PANC-1 and eliminating the need to integrate complex living immune cells. Using the ART-TIME approach, we identify a co-signaling mechanism between PD-1 and other T cell-derived surface receptors as a driver in immune evasion of pancreatic ductal adenocarcinoma. RNA-Seq profiling of WT PANC-1 tumoroids or hybrid tumoroids with integrated synthetic cells carrying various combinations of PD-1 and other surface receptors.