Multiplex Single-Cell Bioprinting for Engineering of Heterogeneous Tissue Constructs with Subcellular Spatial Resolution

Tissue development, function, and disease are largely driven by the spatial organization of individual cells and their interactions within a complex and heterogeneous cellular microenvironment. Engineered tissue models provide a controllable platform to dissect these spatially driven processes. However, strategies to fabricate tissues that replicate near-exact spatial organization and cellular composition of native microenvironments, necessary to faithfully recapitulate in vivo spatial relationships, have remained elusive to date. Here we present a precision bioprinting approach capable of generating high-fidelity replicas of patient biopsies that preserve complex cellular organization and heterogeneity with subcellular resolution. We demonstrate spatial placement of individual cells with down to [~]3 {micro}m spatial precision and up to eight cell types per bioprint. Using this approach, we demonstrate for the first time that altering the microscale spatial organization of tumor microenvironments of similar cellular composition can significantly alter transcriptomic programs of cell response. The ability to engineer and manipulate native cellular microenvironments with single-cell resolution marks a significant advancement toward understanding the critical influence of spatial biology on complex biological systems, such as the tumor microenvironment and organogenesis. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=119 SRC="FIGDIR/small/578499v2_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@11e3f09org.highwire.dtl.DTLVardef@bc7f03org.highwire.dtl.DTLVardef@136acb2org.highwire.dtl.DTLVardef@83e813_HPS_FORMAT_FIGEXP M_FIG C_FIG