Ueyama-Toba Yukiko, Tong Yanran, Yokota Jumpei, Murai Kazuhiro, Hikita Hayato, Eguchi Hidetoshi, Takehara Tetsuo, Mizuguchi Hiroyuki
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Laboratory of Functional Organoid for Drug Discovery, National Institute of Biomedical Innovation, Health and Nutrition, Osaka 567-0085, Japan.
iScience. 2024 Aug 22;27(9):110778. doi: 10.1016/j.isci.2024.110778. eCollection 2024 Sep 20.
Human liver organoids derived from primary human hepatocytes (PHHs) are expected to be a hepatocyte source for preclinical studies of drug metabolism and disposition. Because hepatic functions of these organoids remain low, it is necessary to enhance the hepatic functions. Here, we develop a novel method for two dimensional (2D)-cultured hepatic differentiation from PHH-derived organoids by screening several compounds, cytokines, and growth factors. Hepatic gene expressions in the hepatocyte-like cells differentiated from PHH-derived organoids (Org-HEPs) were greatly increased, compared to those in PHH-derived organoids. The metabolic activities of cytochrome P450 (CYP) 1A2, CYP2C8, CYP2C19, CYP2E1, and CYP3A4 were at levels comparable to those in PHHs. The cell viability of Org-HEPs treated with hepatotoxic drugs was almost the same as that of PHHs. Thus, PHH-derived organoids could be differentiated into highly functional hepatocytes in 2D culture. Thus, Org-HEPs will be useful for pharmaceutical research, including hepatotoxicity tests.
源自原代人肝细胞（PHH）的人肝脏类器官有望成为药物代谢和处置临床前研究的肝细胞来源。由于这些类器官的肝功能仍然较低，因此有必要增强其肝功能。在此，我们通过筛选多种化合物、细胞因子和生长因子，开发了一种从PHH衍生的类器官进行二维（2D）培养的肝分化新方法。与PHH衍生的类器官相比，从PHH衍生的类器官（Org-HEPs）分化而来的肝细胞样细胞中的肝基因表达显著增加。细胞色素P450（CYP）1A2、CYP2C8、CYP2C19、CYP2E1和CYP3A4的代谢活性与PHH中的水平相当。用肝毒性药物处理的Org-HEPs的细胞活力与PHH几乎相同。因此，PHH衍生的类器官可以在二维培养中分化为功能高度健全的肝细胞。因此，Org-HEPs将有助于药物研究，包括肝毒性测试。
