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, scalable and xeno-free integrated differentiation platforms are established to generate hepatic progenitors, mesenchymal stromal cells, and endothelial cells using individual human pluripotent stem cell lines as starting cell types for vascularized liver organoids generation. A scalable microfluidic system is 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 through self-organization. Self-organized vascularized hepatobiliary organoids (VHOs) containing interconnected biliary networks and vascular structures are generated after optimizing the co-culture conditions inside hydrogel microcapsules and transferring the organoids to 3D dynamic suspension culture for further maturation. The VHOs show key functional features 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 can be beneficial to the mass production of human liver organoids for liver organoid medicine and the development of safe, effective, and personalized drugs.

Automated summary

Organoid medicine has made significant progress in the past decade as a novel therapeutic approach for addressing unmet medical needs, such as chronic liver disease. Through the use of scalable and xeno-free differentiation platforms, hepatic progenitors, mesenchymal stromal cells, and endothelial cells are generated from human pluripotent stem cell lines to produce vascularized liver organoids. A scalable microfluidic system is developed to generate cells-loaded microcapsules, which support cell proliferation and the self-organization of liver organoids using a self-biodegradable peptide hydrogel. These vascularized hepatobiliary organoids exhibit key functional features similar to fetal and adult liver tissue, making them promising for liver organoid medicine and drug development.