Heterogeneous liver tissues with multicellular crosstalk and bile duct morphogenesis through organoid bioprinting

Abstract

AbstractLiver is dynamic, heterogeneous, and each cell type acts in concert to regulate its function. Currently, heterogeneous liver tissues are typically built from single cells using bioprinting, making crosstalk between cells difficult. Therefore,in vitromorphogenesis is limited, and self-assembled biliary and blood vessels system are absent from manufactured liver tissues. The combination of bioprinting and organoid technique offers spatial and cellular control over three-dimensional (3D) organ tissue manufacturing, allowing to build liver tissues with self-assembled structurein vitro. We developed a high-throughput PDMS microwell platform (PMP) generating uniform and functional hepatic organoid building blocks (HOBBs) which displayed cellular crosstalk and self-assembled structure. For bioprinting process, we developed three-level temperature control system and new quadratic material, i.e., alginate-gelatin-collagen-laminin (AGCL) biomaterial, realizing reproducible construction of liver tissues with requisite cellular density. Under long-term differentiation, bioprinted liver tissues exhibited enhanced hepatobiliary function, intrahepatic bile duct networks and angiogenic potential. Heterogeneous tissues with coexistence of cholangiocytes, endothelial cells, and hepatocytes was constructed. The heterogeneous liver tissues with angiogenesis and bile duct component (HABs) provides a novel tool for morphogenesis study, liver regeneration, drug testing, and disease research.