Jin Zhe-Long, Xu KangHe, Kim Jonghun, Guo Hao, Yao Xuerui, Xu Yong-Nan, Li Ying-Hua, Ryu DongHee, Kim Kee-Pyo, Hong Kwonho, Kim Yong-June, Wang Lin, Cao Qilong, Kim Kyun-Hwan, Kim Nam-Hyung, Han Dong Wook
Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen, China; International Healthcare Innovation Institute (Jiangmen), Jianghai, Jiangmen, Guangdong Province, China; Research and Development Department, Qingdao Haier Biotech Co. Ltd, Qingdao, China.
Department of Surgery, College of Medicine, Chungbuk National University, Cheongju, 28864, Republic of Korea.
Differentiation. 2024 Jan-Feb;135:100742. doi: 10.1016/j.diff.2023.100742. Epub 2023 Dec 12.
Hepatic organoids might provide a golden opportunity for realizing precision medicine in various hepatic diseases. Previously described hepatic organoid protocols from pluripotent stem cells rely on complicated multiple differentiation steps consisting of both 2D and 3D differentiation procedures. Therefore, the spontaneous formation of hepatic organoids from 2D monolayer culture is associated with a low-throughput production, which might hinder the standardization of hepatic organoid production and hamper the translation of this technology to the clinical or industrial setting. Here we describe the stepwise and fully 3D production of hepatic organoids from human pluripotent stem cells. We optimized every differentiation step by screening for optimal concentrations and timing of differentiation signals in each differentiation step. Hepatic organoids are stably expandable without losing their hepatic functionality. Moreover, upon treatment of drugs with known hepatotoxicity, we found hepatic organoids are more sensitive to drug-induced hepatotoxicity compared with 2D hepatocytes differentiated from PSCs, making them highly suitable for in vitro toxicity screening of drug candidates. The standardized fully 3D protocol described in the current study for producing functional hepatic organoids might serve as a novel platform for the industrial and clinical translation of hepatic organoid technology.
肝脏类器官可能为实现各种肝脏疾病的精准医疗提供一个绝佳机会。先前描述的从多能干细胞生成肝脏类器官的方案依赖于由二维和三维分化程序组成的复杂多步分化过程。因此，从二维单层培养中自发形成肝脏类器官与低通量生产相关，这可能会阻碍肝脏类器官生产的标准化，并妨碍该技术向临床或工业环境的转化。在此，我们描述了从人类多能干细胞逐步且完全三维生成肝脏类器官的方法。我们通过筛选每个分化步骤中分化信号的最佳浓度和时间来优化每个分化步骤。肝脏类器官可稳定扩增且不会丧失其肝脏功能。此外，在用已知肝毒性药物处理后，我们发现与从多能干细胞分化而来的二维肝细胞相比，肝脏类器官对药物诱导的肝毒性更敏感，这使其非常适合用于候选药物的体外毒性筛选。本研究中描述的用于生产功能性肝脏类器官的标准化完全三维方案可能成为肝脏类器官技术工业和临床转化的新平台。
