INSERM Unité Mixte de Recherche (UMR_S) 1193, Villejuif, France; UMR_S 1193, Université Paris-Sud/Paris-Saclay, Villejuif, France; Département Hospitalo-Universitaire Hepatinov, Villejuif, France.
INSERM Unité Mixte de Recherche (UMR_S) 1193, Villejuif, France; UMR_S 1193, Université Paris-Sud/Paris-Saclay, Villejuif, France; Département Hospitalo-Universitaire Hepatinov, Villejuif, France.
Methods Cell Biol. 2020;159:69-93. doi: 10.1016/bs.mcb.2020.03.011. Epub 2020 May 19.
The development of protocols for pluripotent stem cell (PSC) differentiation into cholangiocytes and cholangiocyte organoids in three-dimensional structures represent a huge advance in both research and medical fields because of the limited access to primary human cholangiocytes and the potential bias induced by animal models used to study cholangiopathies in vivo. PSC-derived cholangiocyte organoids consisting of either cysts with luminal space or branching tubular structures are composed of cells with apico-basal polarity that can fulfill cholangiocyte functions like the transport of bile salts. Several protocols of PSC differentiation have already been published but we added to the detailed protocol we describe here some notes or advice to facilitate its handling by new users. We also propose detailed protocols to carry out some of the characterization analyses using immunofluorescence to study the expression of specific markers and a functionality test to visualize bile acid transport using cholyl-lysyl-fluorescein (CLF).
多能干细胞（PSC）分化为胆管细胞和三维结构中的胆管细胞类器官的方案的发展，在研究和医学领域都是一个巨大的进步，因为获得原代人胆管细胞的途径有限，并且动物模型用于体内研究胆管疾病也存在潜在的偏差。PSC 衍生的胆管细胞类器官，由具有腔室的囊泡或分支管状结构组成，由具有顶底极性的细胞组成，能够完成胆汁盐转运等胆管细胞功能。已经发表了几个 PSC 分化的方案，但我们在描述的详细方案中添加了一些注释或建议，以方便新用户使用。我们还提出了详细的方案，使用免疫荧光进行一些特征分析，以研究特定标志物的表达，并使用胆酰-赖氨酰-荧光素（CLF）进行胆汁酸转运的功能测试。
