Laboratory of Stem Cells and Tissue Regeneration, Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, 02841, South Korea.
Department of Predictive Toxicology, Korea Institute of Toxicology, Daejeon, 34114, South Korea.
Biomaterials. 2025 Mar;314:122898. doi: 10.1016/j.biomaterials.2024.122898. Epub 2024 Oct 18.
Hepatic organoids (HOs), primarily composed of hepatobiliary cells, do not represent the pathogenesis of liver diseases due to the lack of non-parenchymal cells. Multi-lineage liver organoids (mLOs) containing various cell types found in the liver offer a promising in vitro disease model. However, their structural complexity remains challenging to achieve due to the difficulty in optimizing culture conditions that meet the growth need of all component cell types. Here, we demonstrate that cystic HOs generated from hPSCs can be expanded long-term and serve as a continuous source for generating complex mLOs. Assembling cystic HOs with hPSC-derived endothelial and hepatic stellate cell-like cells under conventional HO culture conditions failed to support the development of multiple cell types within mLOs, resulting in biased differentiation towards specific cell types. In contrast, modulating the cAMP/Wnt/Hippo signaling pathways with small molecules during assembly and differentiation phases efficiently generate mLOs containing both hepatic parenchymal and non-parenchymal cells. These mLOs exhibited structural complexity and functional maturity, including vascular network formation between parenchymal lobular structures, cell polarity for bile secretion, and the capacity to respond to fibrotic stimuli. Our study underscores the importance of modulating signaling pathways to enhance mLO structural complexity for applications in modeling liver pathologies.
肝类器官（HOs）主要由肝胆细胞组成，由于缺乏非实质细胞，它们不能代表肝脏疾病的发病机制。包含肝脏中各种细胞类型的多谱系肝类器官（mLOs）为体外疾病模型提供了有前途的选择。然而，由于难以优化满足所有组成细胞类型生长需求的培养条件，其结构复杂性仍然难以实现。在这里，我们证明了源自 hPSC 的囊性 HOs 可以长期扩增，并作为生成复杂 mLOs 的连续来源。在传统 HO 培养条件下，将囊性 HOs 与 hPSC 衍生的内皮细胞和肝星状细胞样细胞组装在一起，无法支持 mLO 内多种细胞类型的发育，导致向特定细胞类型的偏分化。相比之下，在组装和分化阶段用小分子调节 cAMP/Wnt/Hippo 信号通路可以有效地生成包含肝实质和非实质细胞的 mLOs。这些 mLOs 表现出结构复杂性和功能成熟性，包括实质小叶结构之间的血管网络形成、胆汁分泌的细胞极性以及对纤维化刺激的反应能力。我们的研究强调了调节信号通路以增强 mLO 结构复杂性在模拟肝脏疾病中的重要性。
