Extrusion bioprinting of cellular aggregates improves mesenchymal stem cell proliferation and differentiation.

3D extrusion bioprinting brings the prospect of stem cell-based therapies in regenerative medicine. These bioprinted stem cells are expected to proliferate and differentiate to form the desired organoids into 3D structures, which is critical for complex tissue construction. However, this strategy is hampered by low reproducible cell number and viability, and organoid immaturity due to incomplete differentiation of stem cells. Hence, we apply a novel extrusion-based bioprinting process with cellular aggregates (CA) bioink, in which the encapsulated cells are precultured in hydrogels to undergo aggregation. In this study, alginate-gelatin-collagen (Alg-Gel-Col) hydrogel containing mesenchymal stem cells (MSCs) were precultured for 48 h to form CA bioink and resulted in high cell viability and printing fidelity. Meanwhile, MSCs in CA bioink showed high proliferation, stemness and lipogenic differentiative potential in contrast to that in single cell (SC) bioink and hanging drop cell spheroid (HDCS) bioink, which indicated the considerable potential for complex tissue construction. In addition, the printability and efficacy of human umbilical cord MSCs (hUC-MSCs) were further confirmed the translational potential of this novel bioprinting method.