Directed differentiation of cholangiocytes from human pluripotent stem cells
Directed differentiation of cholangiocytes from human pluripotent stem cells
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DOI：
10.1038/nbt.3294
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发表时间：
2015-08-01
影响因子：
46.9
通讯作者：
Ghanekar, Anand
中科院分区：
文献类型：
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作者：
Ogawa, Mina;Ogawa, Shinichiro;Ghanekar, Anand
虽然胆管疾病是公认的肝脏疾病的原因，导致胆道功能障碍的分子和细胞事件知之甚少。为了使胆道疾病的建模和药物发现，我们描述了一种方案，该方案通过递送发育相关的线索（包括NOTCH信号传导）来实现胆管上皮细胞（胆管细胞）从人多能干细胞（hPSC）的有效分化。使用三维培养，该方案产生表达成熟胆汁标志物的囊性和/或导管结构，包括顶端钠依赖性胆汁酸转运蛋白、促胰液素受体、纤毛和囊性纤维化跨膜传导调节因子（CFTR）。我们证明，hPSC衍生的胆管细胞具有上皮功能，包括罗丹明流出和CFTR介导的液体分泌。此外，我们表明，功能受损的hPSC衍生的胆管细胞从囊性纤维化患者的CFTR校正拯救。这些发现表明，成熟的胆管细胞可以从hPSC分化，并用于胆道发育和疾病的研究。
Although bile duct disorders are well-recognized causes of liver disease, the molecular and cellular events leading to biliary dysfunction are poorly understood. To enable modeling and drug discovery for biliary disease, we describe a protocol that achieves efficient differentiation of biliary epithelial cells (cholangiocytes) from human pluripotent stem cells (hPSCs) through delivery of developmentally relevant cues, including NOTCH signaling. Using three-dimensional culture, the protocol yields cystic and/or ductal structures that express mature biliary markers, including apical sodium-dependent bile acid transporter, secretin receptor, cilia and cystic fibrosis transmembrane conductance regulator (CFTR). We demonstrate that hPSC-derived cholangiocytes possess epithelial functions, including rhodamine efflux and CFTR-mediated fluid secretion. Furthermore, we show that functionally impaired hPSC-derived cholangiocytes from cystic fibrosis patients are rescued by CFTR correctors. These findings demonstrate that mature cholangiocytes can be differentiated from hPSCs and used for studies of biliary development and disease.