Guan Xiang, Chen Xiaoni, Xu Ying, Wang Lili, Zhang Quan, Luo Xiaoqin, Yin Yunfei, Zhang Ruijia, Liu Xue, Wan Fei, Yang YingCheng, Ge Xiaohu, Wu Di
Department of Developmental and Regenerative Biology, iORGANtech Limited Company (Suzhou), Suzhou, 215000, China.
Department of Central Laboratory, Shenzhen Hospital, Beijing University of Chinese Medicine, Guangdong, 518000, China.
Biochem Biophys Res Commun. 2025 Sep 30;782:152563. doi: 10.1016/j.bbrc.2025.152563. Epub 2025 Aug 30.
Progress in uncovering the causes of extrahepatic biliary diseases and developing new therapies has been constrained by the inaccessibility of donor tissue and a lack of experimental models. Although hepatic, intrahepatic biliary, and pancreatic 2D/3D models have been successfully established from pluripotent stem cells (PSCs), in vitro generation of extrahepatic biliary cells remains a major challenge, due to the absence of developmental cues. Here we report a de novo method for directed differentiation of human PSCs (both embryonic and induced) into pancreato-biliary progenitors-like cells (PBPLCs). We showed that temporal manipulation of the FGF, Vc, WNT, BMP and retinoic acid signaling enabled generation of 3D PBPLCs spheroids manifesting PDX1 SOX17 TBX3 phenotype. These PBPLCs possess bi-potential that could be further directed to Insulin NKX6.1 pancreatic islet β-like cells, or differentiated into gallbladder-like organoids in vitro, respectively, via addition of their developmentally relevant signaling molecules. Formed gallbladder-like organoids are comprised of both epithelial and mesenchymal compartments, self-organized with structural features similar to the native gallbladder. The epithelia in organoids expressed many proteins and genes that highly enriched in extrahepatic cholangiocytes, including SOX17, CLDN3, DBA, CK7, MUC5B, FGF19, CHST4., etc. Our method for generation and purification of PSC-derived PBPLCs opens a new avenue for study of biological mechanisms controlling pancreato-biliary development, as well as their disease modeling and drug screening.
在揭示肝外胆道疾病病因和开发新疗法方面取得的进展一直受到供体组织难以获取和缺乏实验模型的限制。尽管已经成功地从多能干细胞（PSC）建立了肝脏、肝内胆管和胰腺的二维/三维模型，但由于缺乏发育线索，肝外胆管细胞的体外生成仍然是一个重大挑战。在此，我们报告了一种将人PSC（胚胎干细胞和诱导多能干细胞）定向分化为胰腺胆管祖细胞样细胞（PBPLC）的全新方法。我们发现，对FGF、Vc、WNT、BMP和视黄酸信号进行时序调控能够生成表现出PDX1、SOX17、TBX3表型的三维PBPLC球体。这些PBPLC具有双潜能，分别通过添加与其发育相关的信号分子，可进一步定向分化为胰岛素、NKX6.1胰腺胰岛β样细胞，或在体外分化为胆囊样类器官。形成的胆囊样类器官由上皮和间充质部分组成，通过自组织形成与天然胆囊相似的结构特征。类器官中的上皮表达了许多在肝外胆管细胞中高度富集的蛋白质和基因，包括SOX17、CLDN3、DBA、CK7、MUC5B、FGF19、CHST4等。我们生成和纯化PSC来源的PBPLC的方法为研究控制胰腺胆管发育的生物学机制及其疾病建模和药物筛选开辟了一条新途径。
