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

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 (>10). 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.

Bioprinting of Multiscaled Hepatic Lobules within a Highly Vascularized Construct

“…Specifically, they have used iPSC‐derived hepatic endoderm, endothelium, and septum mesenchyme, which enables self‐condensation, hepatic maturation, and vascularization of a successful in vivo transplant. [ 54 ] In this context, we aim to use an iPSC‐derived hepatocyte, which is one of the suitable substitutes of HepG2/C3A, hepatic sinusoidal endothelial cells, hepatic stellate cells, and Kupffer cells or immune cells, as well as primary cells, for hepatic function and tissue‐engineered grafts.…”

A critical look: Challenges in differentiating human pluripotent stem cells into desired cell types and organoids

“…(c) Current-generation hPSC-derived brain organoids possess some key features of early brain development, but various questions remain. hPSC, human pluripotent stem cell hPSC-derived liver precursors and then, at an intermediate step of differentiation, introduced defined numbers of generic endothelial cells (e.g., human umbilical vein endothelial cells) and generic mesenchymal cells to produce threedimensional cultures (Takebe et al, 2013;Takebe et al, 2017). Incorporation of endothelial and mesenchymal cells is an important step to engineer increasingly sophisticated organ simulacra.…”

Optimized protocol for the hepatic differentiation of induced pluripotent stem cells in a fluidic microenvironment

“…Three-dimensional (3D) cultures of liver tissues, based on the spontaneous formation of organoids from a coculture of different cell types, have unequivocally shown that a 3D configuration maintains more efficiently the function and viability of liver cells (Takebe et al, 2017). Indeed, to date, all approaches, including organ decellularization and recellularization, have failed to deliver functional models.…”

“…Indeed, to date, all approaches, including organ decellularization and recellularization, have failed to deliver functional models. Three-dimensional (3D) cultures of liver tissues, based on the spontaneous formation of organoids from a coculture of different cell types, have unequivocally shown that a 3D configuration maintains more efficiently the function and viability of liver cells (Takebe et al, 2017). An alternative liver modeling strategy is to develop both healthy and disease liver-on-chip models.…”