GSE193768
Induced endothelial cell integrated liver assembloids promotes hepatic maturation and therapeutic effect on cholestatic liver fibrosis
Transplantation of pluripotent stem cell (PSC) derived liver organoids have been studied to solve the donor shortage. However, differentiation of unintended cell populations, challenges in generating multi-lineage organoids, and tumorigenicity hinder its uses in clinics. Here, liver assembloids (iHEA) were generated by integrating induced endothelial cells (iEC) and induced hepatic stem cells (iHepSC). iHEA showed enhanced functional maturity iHepSCs derived liver organoids (iHLO) in vitro and had improved therapeutic effects on cholestatic liver fibrosis animals in vivo. Mechanistically, FN1 expressed from iECs led to up-regulation of Itgalph5/beta1 and Hnf4alpha in iHEA and were correlated to decrease of hepatic stellate cells activation genes such as Lox, and Spp1 in the cholestatic liver fibrosis animals. In conclusion, our study demonstrates that directly converted cells can be a promising source for generating transplantable assembloids. This study is designed to generate liver organoids (iHLO) and liver assembloids (iHEA) using induced stem cells generated by direct conversion. Hepatic maturity of iHLO and iHEA were compared by measuring gene expression, albumin/urea secretion and CYP1A2 activity. The experiments were repeated at least three times, and the replicates are indicated in each figure and legend. N values represent the number of animals in the experiment. All animal experimental procedures were approved and conducted under a protocol (UNISTIACUC-20-49) by the Animal Care and Use Committee of Ulsan National Institute of Science and Technology (UNIST, Ulsan, South Korea). The experiments were carried out in accordance with documented standards of the Institutional Review Board of Ulsan National Institute of Science and Technology (UNIST) (UNISTIRB-15–17 C) for human cell experiments. Overall design: The data Fib (Fibroblast) is performed with 1 replicate The data 2D-iHepSC (2 Dimensional induced Hepatic Stem Cells (2D-iHepSCs)) is performed with 1 replicate The data 2D-iHepSCiEC (2 Dimensional induced Hepatic Stem Cells integrating induced endothelial cells (2D-iHepSCECs)) is performed with 1 replicate The data 3D-iHA (3 Dimensional Hepatic Assembloids (3D-iHAs)) is performed with 1 replicate The data 3D-iHEA (3 Dimensional Hepatic Endothelial Assembloids (3D-iHEAs)) is performed with 1 replicate The data Adult-Liver (Adult-Liver1) is performed with 1 replicate The data Fetal-Liver (Fetal-Liver1) is performed with 1 replicate