3D Bioprinting Lobule-like Hepatorganoids with Induced Vascularization for Orthotopic Implantation

Orthotopic implantation tumor assays are frequently employed in tumor biology, drug screening, and personalized therapy researches. However, the limited representation of a complex tumor microenvironment in these implanted animal models hampers a comprehensive understanding of tumor progression and therapeutic response. In this research, we developed a 3D bioprinting method using gelatin methacryloyl (GelMA) hydrogel loaded with a prolonged released system of vascular endothelial growth factor (VEGF), embedding hepatocellular carcinoma cells, to fabricate lobule-like hepatorganoids that faithfully mimic the architecture of hepatic lobules. Moreover, we utilized GelMA hydrogel mixed with Matrigel, containing human umbilical vein endothelial cells (HUVECs), to induce vascularization on the organoids. Orthotopic implantation in mouse model was performed, allowing for an improved simulation of the vascularization process within hepatorganoids. Compared to control group, 3D bioprinted lobule-like hepatorganoids exhibited prolonged survival in mice, while serum analysis revealed significantly elevated levels of alpha-fetoprotein (AFP). Eventually, our findings demonstrate that this system effectively forms orthotopic implantion of liver tumors and facilitates vascularization, which may notably contribute to the understanding of tumor biology, drug screening, and personalized therapy. Overall design: Gene expression profiles of 3D bioprinted vascularized lobule-like hepatorganoids (VLH), 3D bioprinted HUVECs-hepatorganoids (HH), 3D bioprinted VEGF/hepatorganoids (VH), 3D bioprinted hepatorganoids (3DC) and 2D cultured huh7 (2DC) cells.