Human branching cholangiocyte organoids recapitulate embryogenic bile duct development and provide an unique model to study biliary diseases. scRNA-Seq. Human branching cholangiocyte organoids recapitulate embryogenic bile duct development and provide an unique model to study biliary diseases. scRNA-Seq

Although providing promising and unique tools for studying cholangiocytes, current tissue-derived cholangiocyte-organoid systems do not recapitulate the complex architecture of the intrahepatic bile ducts in vitro. Here, we report a new method for creating branching cholangiocyte organoids (BRCO) from human adult tissue to study the intrahepatic biliary tree and diseases. BRCOs self-organize into large complex tubular structures, while closely resembling primary cholangiocytes on a transcriptomic and functional level. They are capable of mimicking branching bile duct development as well as being used for studying diseases in which the biliary tree does not develop properly (Alagille Syndrome). Furthermore, we deliver evidence that our culture method allows for formation of complex cholangiocyte cancer (cholangiocarcinoma, CCA) organoids. These branching CCA organoids resemble the primary tumor more closely compared to previously published protocols as well as showing unique tumor-specific drug responses. In conclusion, our culture method allows for creation of novel (malignant) cholangiocyte-organoids to study the intrahepatic bile ducts. Overall design: single cell transcriptomic profiles of ICO and BRCO samples

尽管现有组织来源的胆管上皮细胞类器官（cholangiocyte-organoid）系统可为胆管上皮细胞（cholangiocyte）的研究提供颇具前景且独特的研究工具，但这类系统无法在体外重现肝内胆管（intrahepatic bile ducts）的复杂解剖结构。本研究报道了一种全新方法，可从成人人体组织中构建分支状胆管上皮细胞类器官（branching cholangiocyte organoids, BRCO），用于肝内胆道系统及其相关疾病的研究。BRCO可自组装形成大型复杂管状结构，且在转录组与功能层面与原代胆管上皮细胞高度相似。该类器官能够模拟分支状胆管的发育过程，同时可用于研究胆道系统发育异常的疾病（如阿拉基综合征（Alagille Syndrome））。此外，本研究证实该培养方法可构建复杂的胆管癌（cholangiocarcinoma, CCA）类器官。与已发表的现有培养方案相比，此类分支状CCA类器官与原发肿瘤的匹配度更高，且展现出独特的肿瘤特异性药物响应特征。综上，本研究的培养方法可构建新型（恶性）胆管上皮细胞类器官，用于肝内胆管的相关研究。整体实验设计：ICO与BRCO样本的单细胞转录组图谱。