Erasmus MC Transplant Institute, University Medical Center Rotterdam, Department of Surgery, Rotterdam, the Netherlands; Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, UK.
Erasmus MC Transplant Institute, University Medical Center Rotterdam, Department of Surgery, Rotterdam, the Netherlands.
Cell Stem Cell. 2022 May 5;29(5):776-794.e13. doi: 10.1016/j.stem.2022.04.011.
Human cholangiocyte organoids show great promise for regenerative therapies and in vitro modeling of bile duct development and diseases. However, the cystic organoids lack the branching morphology of intrahepatic bile ducts (IHBDs). Here, we report establishing human branching cholangiocyte organoid (BRCO) cultures. BRCOs self-organize into complex tubular structures resembling the IHBD architecture. Single-cell transcriptomics and functional analysis showed high similarity to primary cholangiocytes, and importantly, the branching growth mimics aspects of tubular development and is dependent on JAG1/NOTCH2 signaling. When applied to cholangiocarcinoma tumor organoids, the morphology changes to an in vitro morphology like primary tumors. Moreover, these branching cholangiocarcinoma organoids (BRCCAOs) better match the transcriptomic profile of primary tumors and showed increased chemoresistance to gemcitabine and cisplatin. In conclusion, BRCOs recapitulate a complex process of branching morphogenesis in vitro. This provides an improved model to study tubular formation, bile duct functionality, and associated biliary diseases.
人胆管细胞类器官在再生疗法和胆管发育及疾病的体外模型方面具有巨大的应用潜力。然而，这些囊状类器官缺乏肝内胆管（IHBD）的分支形态。在这里，我们报告了建立人分支胆管细胞类器官（BRCO）培养物的方法。BRCO 能够自我组织成复杂的管状结构，类似于 IHBD 结构。单细胞转录组学和功能分析表明，它与人原代胆管细胞高度相似，而且重要的是，其分支生长模拟了管状发育的某些方面，并且依赖于 JAG1/NOTCH2 信号通路。当将其应用于胆管癌肿瘤类器官时，其形态会发生变化，呈现出类似于原发性肿瘤的体外形态。此外，这些分支胆管癌细胞类器官（BRCCAOs）与原发性肿瘤的转录组谱更为匹配，并表现出对吉西他滨和顺铂的化疗耐药性增加。总之，BRCO 能够在体外重现分支形态发生的复杂过程。这为研究管状形成、胆管功能和相关胆道疾病提供了一个改进的模型。
