Fan Han, Shang Jia, Li Junbo, Yang Bo, Zhou Ding, Jiang Shanqing, Fan Yuhang, Zhou Ying, Wang Yuwen, Liu Peidi, Li Changyong, Chen Zhishui, Chen Pu
Tissue Engineering and Organ Manufacturing (TEOM) Lab, Department of Biomedical Engineering, Wuhan University TaiKang Medical School (School of Basic Medical Sciences), Wuhan, 430071, China.
Department of Biological Repositories, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China.
Adv Sci (Weinh). 2025 Feb;12(8):e2407945. doi: 10.1002/advs.202407945. Epub 2025 Jan 4.
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.
肝脏类器官已越来越多地被用作研究肝脏发育和疾病的关键体外模型。然而，在不影响肝脏实质细胞分化的情况下实现肝脏类器官的血管预构建仍然是一个长期挑战，这对于其在再生医学中的应用至关重要。在此，通过一种新策略，即非实质细胞移植（NCG），在不影响肝脏上皮细胞分化的情况下实现了预血管化人肝胆类器官（vhHBOs）的大规模形成。在内皮细胞和间充质细胞处于不同肝脏上皮分化阶段时，将它们移植到人类肝胆类器官（hHBOs）中，而不添加非实质细胞培养基和生长因子。与肝脏细胞分化阶段相比，在肝脏成熟阶段进行内皮细胞移植可提供最佳整合效率。此外，在vhHBOs构建过程中，间充质细胞移植对于内皮细胞侵入并向肝上皮细胞发芽至关重要。将异位肝脏植入小鼠体内进一步显示vhHBOs整合到小鼠血管网络中。值得注意的是，移植的vhHBOs会自我组织成类似肝小叶和胆管的天然肝组织，表明它们具有再生受损肝组织和胆管组织的潜力。据信，非实质细胞移植将为组织工程和再生医学提供一种新的技术途径，以形成高保真的体外复杂模型。
