Electrostatically Cross-Linked Bioinks for Jetting-Based Bioprinting of 3D Cell Cultures

It has been acknowledged that thousands of drugs that passed two-dimensional (2D) cell culture models and animal studies often fail when entering human clinical trials. Despite the significant development of three-dimensional (3D) models, developing a high-throughput model that can be reproducible on a scale remains challenging. One of the main challenges is precise cell deposition and the formation of a controllable number of spheroids to achieve more reproducible results for drug discovery and treatment applications. Furthermore, when transitioning from manually generated structures to 3D bioprinted structures, the choice of material is limited due to restrictions on materials that are applicable with bioprinters. Herein, we have shown the capability of a fast-cross-linking bioink that can be used to create a single spheroid with varying diameters (660, 1100, and 1340 μm) in a high-throughput manner using a commercialized drop-on-demand bioprinter. Throughout this work, we evaluate the physical properties of printable ink with and without cells, printing optimization, cytocompatibility, cell sedimentation, and homogeneity in ink during the printing process. This work showcases the importance of ink characterization to determine printability and precise cell deposition. The knowledge gained from this work will accelerate the development of next-generation inks compatible with a drop-on-demand 3D bioprinter for various applications such as precision models to mimic diseases, toxicity tests, and the drug development process.