Self-Assembled Matrigel-Free iPSC-Derived Liver Organoids Demonstrate Wide-Ranging Highly Differentiated Liver Functions

Human induced pluripotent stem cell (iPSC)-derived liver organoids serve as models of organogenesis, disease, drug screening, and regenerative medicine. Prevailing methods for generating organoids rely on Matrigel, whose batch-to-batch variability and xenogeneic source pose challenges to mechanistic research and translation to human clinical therapy. In this report, we demonstrate that self-assembled Matrigel-free iPSC-derived organoids developed in rotating wall vessels (RWVs) exhibit greater hepatocyte-specific functions than organoids formed on Matrigel. We show that RWVs produce highly functional liver organoids in part by eliminating the need for Matrigel, which has adverse effects on hepatic lineage differentiation. RWV liver organoids sustain durable function over long-term culture and express a range of mature functional genes at levels comparable to adult human liver, while retaining some fetal features. Our results indicate that RWVs provide a simple and high-throughput way to generate Matrigel-free liver organoids suitable for research and clinical applications. Organoid self-assembly within rotating wall vessels (RWV) is a simple and high-throughput way to generate highly functional human-induced pluripotent stem cell-derived liver organoids without the need for Matrigel. Liver organoids self-assembled in RWVs demonstrate higher, broader, and more durable hepatic function than organoids generated on Matrigel, in part because Matrigel has some adverse effects on hepatic lineage commitment.

Automated summary

Organoid self-assembly within rotating wall vessels (RWV) is a simple and high-throughput way to generate highly functional human-induced pluripotent stem cell-derived liver organoids without the need for Matrigel. Liver organoids self-assembled in RWVs demonstrate higher, broader, and more durable hepatic function than organoids generated on Matrigel, in part because Matrigel has some adverse effects on hepatic lineage commitment.