Patterning of Artificial Cell Populations Via Laser Induced Forward Transfer for Controlled and Localized Cargo Uptake and Release

Complex coacervate-based synthetic cells are widely used to mimic biologically relevant cellular processes, most often from the single-cell perspective. However, in nature, multicellular organization is a crucial driver of function. Synthetic cell research, therefore, requires fabrication methods that allow control over the spatial organization of synthetic cells to achieve translational potential. Here, we developed an approach to position polymer-stabilized complex coacervates within biopolymer matrices in an adherent manner. To accurately deliver and pattern the coacervates onto tissue culture substrates, we investigated laser-induced forward transfer (LIFT) 3D printing and screened printing parameters, including laser energy and the number of stacked layers, to optimize the printability of the complex coacervate-adhered biomaterial ink. We demonstrated precise spatial control over coacervate localization at the population level by co-patterning multiple coacervate populations into a biomaterial matrix. Lastly, we showed that both the adhered complex-coacervation method and LIFT printing process preserve basic biologically relevant functionality, as demonstrated by protein cargo uptake into and release from the patterned synthetic cells.