Cell printing has found wide applications in biomedical fields due to its unique capability in fabricating living tissue constructs with precise control over cell arrangements. However, it is still challenging to print cell-laden 3D structures simultaneously with high resolution and high cell viability. Here a coaxial nozzle-assisted electrohydrodynamic cell printing strategy was developed to fabricate living 3D cell-laden constructs. Critical process parameters such as feeding rate and stage moving speed were evaluated to achieve smaller hydrogel filaments. The effect of CaCl 2 feeding rate on the printing of 3D alginate hydrogel constructs was also investigated. The results indicated that the presented strategy can print 3D hydrogel structures with relatively uniform filament dimension (about 80 μm) and cell distribution. The viability of the encapsulated cells was over 90%. We envision that the coaxial nozzle-assisted electrohydrodynamic printing will become a promising cell printing strategy to advance biomedical innovations. Keywords: electrohydrodynamic printing; cell printing; bioprinting; biofabrication; tissue engineering

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Recapitulating the vascular networks that maintain the delivery of nutrition, oxygen, and byproducts for the living cells within the three-dimensional (3D) tissue constructs is a challenging issue in the tissue-engineering area. Here, a novel coaxial electrohydrodynamic (EHD) bioprinting strategy is presented to fabricate thick pre-vascularized cell-laden constructs. The alginate and collagen/calcium chloride solution were utilized as the outer-layer and inner-layer bioink, respectively, in the coaxial printing nozzle to produce the core-sheath hydrogel filaments. The effect of process parameters (the feeding rate of alginate and collagen and the moving speed of the printing stage) on the size of core and sheath lines within the printed filaments was investigated. The core-sheath filaments were printed in the predefined pattern to fabricate lattice hydrogel with perfusable lumen structures. Endothelialized lumen structures were fabricated by culturing the core-sheath filaments with endothelial cells laden in the core collagen hydrogel. Multilayer core-sheath filaments were successfully printed into 3D porous hydrogel constructs with a thickness of more than 3 mm. Finally, 3D pre-vascularized cardiac constructs were successfully generated, indicating the efficacy of our strategy to engineer living tissues with complex vascular structures.