Massive and Reproducible Production of Liver Buds Entirely from Human Pluripotent Stem Cells

TLDR

Organoid technology offers a revolutionary therapeutic paradigm, yet its human application has been limited by reproducibility and scalability challenges. This study develops a scalable, reproducible platform for producing liver buds entirely from human iPSCs. By reverse‑screening, we identified hepatic endoderm, endothelial, and septum mesenchyme progenitors that reliably generate liver buds, and we scaled production using an omni‑well array capable of >10⁸ homogeneous miniaturized buds. The resulting vascularized, functional iPSC‑derived liver tissues enhance hepatic function through stage‑matched progenitor interactions and rescue acute liver failure upon transplantation, establishing a robust manufacturing platform for clinical and pharmaceutical use.

Abstract

Organoid technology provides a revolutionary paradigm toward therapy but has yet to be applied in humans, mainly because of reproducibility and scalability challenges. Here, we overcome these limitations by evolving a scalable organ bud production platform entirely from human induced pluripotent stem cells (iPSC). By conducting massive "reverse" screen experiments, we identified three progenitor populations that can effectively generate liver buds in a highly reproducible manner: hepatic endoderm, endothelium, and septum mesenchyme. Furthermore, we achieved human scalability by developing an omni-well-array culture platform for mass producing homogeneous and miniaturized liver buds on a clinically relevant large scale (>108). Vascularized and functional liver tissues generated entirely from iPSCs significantly improved subsequent hepatic functionalization potentiated by stage-matched developmental progenitor interactions, enabling functional rescue against acute liver failure via transplantation. Overall, our study provides a stringent manufacturing platform for multicellular organoid supply, thus facilitating clinical and pharmaceutical applications especially for the treatment of liver diseases through multi-industrial collaborations.