Continuous Production of Highly Functional Vascularized Hepatobiliary Organoids from Human Pluripotent Stem Cells using a Scalable Microfluidic Platform.

"Organoid medicine" has rapidly progressed over the past decade as a new class of therapeutics with high functionality and complexity for addressing unmet medical needs such as effective treatment of patients suffering from chronic liver disease using liver organoids. Here, we established scalable and xeno-free integrated differentiation platforms to generate hepatic progenitors, mesenchymal stromal cells, and endothelial cells using individual human pluripotent stem cell lines (hPSCs) as starting cell types for vascularized liver organoids generation. A scalable microfluidic system was developed to continuously generate cells-loaded microcapsules with self-biodegradable 4-arm-PEG-MMP1-sensitive peptide hydrogel as shell material, to support cells proliferation, self-condensation, and liver organoids generation thorough self-organization. Self-organized vascularized hepatobiliary organoids containing inter-connected biliary networks and vascular structures were generated after optimizing the co-culture conditions inside hydrogel microcapsules and transferring the organoids to 3D dynamic suspension culture for further maturation. The vascularized hepatobiliary organoids showed key functional features that were similar to the fetal and adult liver tissue including the expression of liver-specific marker genes, the ability to perform main liver metabolic functions, and inducing drug metabolism. The established platforms could be beneficial to the mass production of human liver organoids for liver organoid medicine and the development of safe, effective, and personalized drugs.