High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell Grafting

Abstract Liver organoids have been increasingly adopted as a critical in vitro model to study liver development and diseases. However, the pre‐vascularization of liver organoids without affecting liver parenchymal specification remains a long‐lasting challenge, which is essential for their applicati...

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Main Authors: Han Fan, Jia Shang, Junbo Li, Bo Yang, Ding Zhou, Shanqing Jiang, Yuhang Fan, Ying Zhou, Yuwen Wang, Peidi Liu, Changyong Li, Zhishui Chen, Pu Chen
Format: Article
Language:English
Published: Wiley 2025-02-01
Series:Advanced Science
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Online Access:https://doi.org/10.1002/advs.202407945
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author Han Fan
Jia Shang
Junbo Li
Bo Yang
Ding Zhou
Shanqing Jiang
Yuhang Fan
Ying Zhou
Yuwen Wang
Peidi Liu
Changyong Li
Zhishui Chen
Pu Chen
author_facet Han Fan
Jia Shang
Junbo Li
Bo Yang
Ding Zhou
Shanqing Jiang
Yuhang Fan
Ying Zhou
Yuwen Wang
Peidi Liu
Changyong Li
Zhishui Chen
Pu Chen
author_sort Han Fan
collection DOAJ
description Abstract Liver organoids have been increasingly adopted as a critical in vitro model to study liver development and diseases. However, the pre‐vascularization of liver organoids without affecting liver parenchymal specification remains a long‐lasting challenge, which is essential for their application in regenerative medicine. Here, the large‐scale formation of pre‐vascularized human hepatobiliary organoids (vhHBOs) is presented without affecting liver epithelial specification via a novel strategy, namely nonparenchymal cell grafting (NCG). Endothelial and mesenchymal cells are grafted to human hepatobiliary organoids (hHBOs) at the different liver epithelial differentiation stages without supplementing with nonparenchymal culture medium and growth factors. Endothelial grafting at the stage of hepatic maturation offers an optimal integration efficiency compared to the stage of hepatic specification. Additionally, grafting with mesenchymal proves crucial in endothelial invading and sprouting into the liver epithelial cells during the establishment of vhHBOs. Ectopic liver implants into mice further displayed integration of vhHBOs into mice vascular networks. Notably, transplanted vhHBOs self‐organized into native liver tissue like hepatic zone and bile ducts, indicating their potential to regenerate damaged hepatic and bile duct tissues. It is believed that nonparenchymal cell grafting will offer a novel technical route to form a high‐fidelity complex in vitro model for tissue engineering and regenerative medicine.
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spelling doaj-art-0d6a8895cb264b119878bb991e96ff602025-08-20T02:30:35ZengWileyAdvanced Science2198-38442025-02-01128n/an/a10.1002/advs.202407945High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell GraftingHan Fan0Jia Shang1Junbo Li2Bo Yang3Ding Zhou4Shanqing Jiang5Yuhang Fan6Ying Zhou7Yuwen Wang8Peidi Liu9Changyong Li10Zhishui Chen11Pu Chen12Tissue Engineering and Organ Manufacturing (TEOM) Lab Department of Biomedical Engineering Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan 430071 ChinaDepartment of Biological Repositories Zhongnan Hospital of Wuhan University Wuhan 430071 ChinaKey Laboratory of Organ Transplantation Institute of Organ Transplantation Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 ChinaKey Laboratory of Organ Transplantation Institute of Organ Transplantation Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 ChinaTissue Engineering and Organ Manufacturing (TEOM) Lab Department of Biomedical Engineering Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan 430071 ChinaTissue Engineering and Organ Manufacturing (TEOM) Lab Department of Biomedical Engineering Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan 430071 ChinaTissue Engineering and Organ Manufacturing (TEOM) Lab Department of Biomedical Engineering Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan 430071 ChinaResearch Center for Medicine and Structural Biology of Wuhan University Wuhan University Wuhan Hubei 430071 ChinaTissue Engineering and Organ Manufacturing (TEOM) Lab Department of Biomedical Engineering Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan 430071 ChinaTissue Engineering and Organ Manufacturing (TEOM) Lab Department of Biomedical Engineering Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan 430071 ChinaDepartment of Physiology Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan Hubei 430071 ChinaKey Laboratory of Organ Transplantation Institute of Organ Transplantation Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 ChinaTissue Engineering and Organ Manufacturing (TEOM) Lab Department of Biomedical Engineering Wuhan University TaiKang Medical School (School of Basic Medical Sciences) Wuhan 430071 ChinaAbstract Liver organoids have been increasingly adopted as a critical in vitro model to study liver development and diseases. However, the pre‐vascularization of liver organoids without affecting liver parenchymal specification remains a long‐lasting challenge, which is essential for their application in regenerative medicine. Here, the large‐scale formation of pre‐vascularized human hepatobiliary organoids (vhHBOs) is presented without affecting liver epithelial specification via a novel strategy, namely nonparenchymal cell grafting (NCG). Endothelial and mesenchymal cells are grafted to human hepatobiliary organoids (hHBOs) at the different liver epithelial differentiation stages without supplementing with nonparenchymal culture medium and growth factors. Endothelial grafting at the stage of hepatic maturation offers an optimal integration efficiency compared to the stage of hepatic specification. Additionally, grafting with mesenchymal proves crucial in endothelial invading and sprouting into the liver epithelial cells during the establishment of vhHBOs. Ectopic liver implants into mice further displayed integration of vhHBOs into mice vascular networks. Notably, transplanted vhHBOs self‐organized into native liver tissue like hepatic zone and bile ducts, indicating their potential to regenerate damaged hepatic and bile duct tissues. It is believed that nonparenchymal cell grafting will offer a novel technical route to form a high‐fidelity complex in vitro model for tissue engineering and regenerative medicine.https://doi.org/10.1002/advs.202407945hepatobiliary organoids on a chipmicrofabricated hexagonal closely packed cavity arraysnonparenchymal cell graftingpre‐vascularization
spellingShingle Han Fan
Jia Shang
Junbo Li
Bo Yang
Ding Zhou
Shanqing Jiang
Yuhang Fan
Ying Zhou
Yuwen Wang
Peidi Liu
Changyong Li
Zhishui Chen
Pu Chen
High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell Grafting
Advanced Science
hepatobiliary organoids on a chip
microfabricated hexagonal closely packed cavity arrays
nonparenchymal cell grafting
pre‐vascularization
title High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell Grafting
title_full High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell Grafting
title_fullStr High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell Grafting
title_full_unstemmed High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell Grafting
title_short High‐Throughput Formation of Pre‐Vascularized hiPSC‐Derived Hepatobiliary Organoids on a Chip via Nonparenchymal Cell Grafting
title_sort high throughput formation of pre vascularized hipsc derived hepatobiliary organoids on a chip via nonparenchymal cell grafting
topic hepatobiliary organoids on a chip
microfabricated hexagonal closely packed cavity arrays
nonparenchymal cell grafting
pre‐vascularization
url https://doi.org/10.1002/advs.202407945
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