Data_Sheet_1_Human Bile Contains Cholangiocyte Organoid-Initiating Cells Which Expand as Functional Cholangiocytes in Non-canonical Wnt Stimulating Conditions.PDF

Diseases of the bile duct (cholangiopathies) remain a common indication for liver transplantation, while little progress has been made over the last decade in understanding the underlying pathophysiology. This is largely due to lack of proper in vitro model systems to study cholangiopathies. Recently, a culture method has been developed that allows for expansion of human bile duct epithelial cells grown as extrahepatic cholangiocyte organoids (ncECOs) in non-canonical Wnt-stimulating conditions. These ncECOs closely resemble cholangiocytes in culture and have shown to efficiently repopulate collagen scaffolds that could act as functional biliary tissue in mice. Thus far, initiation of ncECOs required tissue samples, thereby limiting broad patient-specific applications. Here, we report that bile fluid, which can be less invasively obtained and with low risk for the patients, is an alternative source for culturing ncECOs. Further characterization showed that bile-derived cholangiocyte organoids (ncBCOs) are highly similar to ncECOs obtained from bile duct tissue biopsies. Compared to the previously reported bile-cholangiocyte organoids cultured in canonical Wnt-stimulation conditions, ncBCOs have superior function of cholangiocyte ion channels and are able to respond to secretin and somatostatin. In conclusion, bile is a new, less invasive, source for patient-derived cholangiocyte organoids and makes their regenerative medicine applications more safe and feasible.

胆管疾病（胆管病理学）已成为肝移植的常见指征，然而在过去十年中，对潜在病理生理学的理解进展有限。这主要归因于缺乏适当的体外模型系统来研究胆管病理学。近期，一种培养方法被开发出来，该方法允许在非经典Wnt刺激条件下，将培养的人胆管上皮细胞扩展为胆管细胞器官oids（ncECOs）。这些ncECOs在培养中与胆管细胞相似，并显示出能够高效地再充填胶原蛋白支架，这些支架在老鼠中可以作为功能性胆管组织。迄今为止，ncECOs的启动需要组织样本，从而限制了广泛的患者特异性应用。在此，我们报告称，胆汁，其可以通过更少的侵入性获得且对患者风险较低，是培养ncECOs的替代来源。进一步的表征表明，源自胆汁的胆管细胞器官oids（ncBCOs）与来自胆管组织活检获得的ncECOs高度相似。与之前报道的在经典Wnt刺激条件下培养的胆汁-胆管细胞器官oids相比，ncBCOs在胆管细胞离子通道功能方面表现出优越性，并能对促胰液素和生长抑素做出反应。总之，胆汁是患者来源的胆管细胞器官oids的新来源，且更少侵入性，使其再生医学应用更为安全可行。