Tong Yanran, Ueyama-Toba Yukiko, 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.
Biochem Biophys Rep. 2023 Jan 21;33:101432. doi: 10.1016/j.bbrep.2023.101432. eCollection 2023 Mar.
Organoid culture is a technology for creating three-dimensional (3D) tissue-like structures , and is expected to be used in various fields. It was reported that human adult bile duct cells derived from human biopsy can be expanded as organoids that exhibit stem cell-like properties including high proliferative ability and differentiation ability toward both hepatocytes and biliary epithelial cells (BECs). Although many studies have achieved the efficient differentiation of bipotent human liver-derived organoids (hLOs) toward mature hepatocytes, the differentiation potency toward mature BECs remains unclear. In this study, we attempted to evaluate the differentiation potency of bipotent hLOs, which were generated from primary (cryopreserved) human hepatocytes (PHHs), toward BECs by sequential treatment with epidermal growth factor (EGF), Interleukin-6 (IL-6), and sodium taurocholate hydrate. Along with the differentiation toward bipotent hLOs-derived BECs (Org-BECs), increases in the gene expression levels of BEC markers and formation of the lumen-like structures typical of BECs were observed. In addition, Org-BECs exhibited P-glycoprotein-mediated drug transport capacity. Finally, in order to expand the applicability of Org-BECs, we succeeded in the differentiation of bipotent hLOs toward BECs in a two-dimensional (2D) culture system. Our findings demonstrated that bipotent hLOs can indeed differentiate into mature BECs, meaning that they possess a capacity for differentiation toward both hepatocytes and BECs.
类器官培养是一种用于创建三维（3D）组织样结构的技术，有望应用于各个领域。据报道，源自人类活检的成人胆管细胞可作为类器官进行扩增，这些类器官表现出干细胞样特性，包括高增殖能力以及向肝细胞和胆管上皮细胞（BEC）分化的能力。尽管许多研究已实现双能性人肝源性类器官（hLO）向成熟肝细胞的高效分化，但向成熟BEC的分化潜能仍不清楚。在本研究中，我们试图通过用表皮生长因子（EGF）、白细胞介素-6（IL-6）和牛磺胆酸钠水合物进行序贯处理，评估由原代（冷冻保存）人肝细胞（PHH）生成的双能性hLO向BEC的分化潜能。随着向双能性hLO衍生的BEC（Org-BEC）的分化，观察到BEC标志物基因表达水平的增加以及BEC典型的管腔样结构的形成。此外，Org-BEC表现出P-糖蛋白介导的药物转运能力。最后，为了扩大Org-BEC的适用性，我们成功地在二维（2D）培养系统中将双能性hLO分化为BEC。我们的研究结果表明，双能性hLO确实可以分化为成熟的BEC，这意味着它们具有向肝细胞和BEC分化的能力。
