Bioprinting 3D muscle tissue supplemented with endothelial-spheroids for neuromuscular junction model

The neuromuscular junction (NMJ) plays a critical role in muscle contraction, and its dysfunction can result various disorders. In vitro models for studying NMJ are essential understanding their functions pathology. However, the engineering of tissue presents challenges organization myofiber-like oriented bundles as well induction vessel formation innervation. To address these challenges, we fabricated hybrid construct comprising uniaxially aligned struts endothelial cell spheroids using combination situ electric field-assisted bioprinting (E-printing) microdroplet-based spheroid-forming (MDS-printing) techniques. This resulted self-aggregation human umbilical vein cells (HUVECs) into without attachment to structures. We tested fabrication parameters, such field cross-linking conditions, E-printing deposited density MDS-printing, stabilize alignment progenitor (hMPCs) HUVEC spheroids, respectively. stimulated hMPCs efficiently formed fully myofibers, incorporation induced highly upregulated crosstalk between different types compared simple E-printed hMPC/HUVEC mixture-loaded construct. improved myogenesis vitro. addition, when co-cultured with motor neuron-like (NSC-34) spheroid separated by channel, observed considerably those normally mixed cell-bearing Our findings suggest that this has potential enhance improving biological activities through HUVEC-spheroids facilitating co-culture NSC-34 spheroids.