Generation of expandable hepatocyte organoids with multipolar organization from human embryonic stem cells

In this study, we successfully generated mature grape-like heporgs (G-heporg) from human embryonic stem cells (hESCs) using a modified hepatocyte organoid culture system. Within this differentiation platform, spherical hepatocyte organoids (S-heporg) spontaneously emerged alongside G-heporgs. Similar to primary heporgs, differentiated G-heporgs exhibited high ALB expression and expansion capability, whereas the S-heporgs lacked both characteristics. Comparative analysis revealed that IGF2 stimulates G-heporg formation. Moreover, we found that targeted modulation of the Hippo-YAP pathway overcame long-term expansion limitations in G-heporgs. Crucially, the differentiated and expandable G-heporgs developed multipolar architectures with functional BC networks, recapitulating the polarity characteristic of primary hepatocytes. Finally, we modeled copper ion metabolism using G-heporgs, demonstrating their utility for investigating polarized trafficking processes of hepatocyte metabolites