Efficient and Controlled Generation of 2D and 3D Bile Duct Tissue from Human Pluripotent Stem Cell-Derived Spheroids.
Efficient and Controlled Generation of 2D and 3D Bile Duct Tissue from Human Pluripotent Stem Cell-Derived Spheroids.
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DOI：
10.1007/s12015-016-9657-5
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发表时间：
2016-08
影响因子：
4.8
通讯作者：
Jang YY
中科院分区：
文献类型：
--
作者：
Tian L;Deshmukh A;Ye Z;Jang YY
在过去的几年里，体外肝组织工程的研究越来越多，但目前的工程肝组织缺乏胆管系统。胆汁引流不足不仅会阻碍肝脏的基本消化功能，还会导致胆汁的积累，而胆汁对肝细胞有毒，已知会导致肝硬化。显然，胆管组织的生成对于肝脏的工程功能和健康至关重要。人诱导多能干细胞（iPSCs）分化为胆管组织需要长时间和/或复杂的培养条件，并且迄今为止效率低下。为了生成一个功能齐全的含有胆道系统的肝脏，我们已经开发了有效的2D和3D胆管上皮组织生成的定义和控制条件。成人肝脏和人胎儿肝脏导管板中多能肝祖细胞的标记EpCAM在人多能干细胞生成的肝球体中高度表达。EpCAM高肝球不仅能在二维分化条件下高效生成单层胆道上皮细胞，还能在三维条件下形成功能性的导管结构。重要的是，这种EpCAM高球型胆道组织生成比其他现有方法要快得多，而且不需要细胞分选。此外，我们发现通过crispr /Cas9基因组编辑技术产生的敲入CK7报告基因的人类iPSC系极大地促进了胆道分化的分析。这种新的胆管分化方法将为获得胆管细胞和组织提供一种更有效的方法，为将来完整和功能性肝组织的工程化提供便利。
While in vitro liver tissue engineering has been increasingly studied during the last several years, presently engineered liver tissues lack the bile duct system. The lack of bile drainage not only hinders essential digestive functions of the liver, but also leads to accumulation of bile that is toxic to hepatocytes and known to cause liver cirrhosis. Clearly, bile duct tissue generation is essential for engineering functional and healthy liver. Differentiation of human induced pluripotent stem cells (iPSCs) to bile duct tissue requires long and/or complex culture conditions, and has been inefficient so far. Towards generating a fully functional liver containing biliary system, we have developed defined and controlled conditions for efficient 2D and 3D bile duct epithelial tissue generation. A marker for multipotent liver progenitor in both adult human liver and ductal plate in human fetal liver, EpCAM, is highly expressed in hepatic spheroids generated from human iPSCs. The EpCAM high hepatic spheroids can, not only efficiently generate a monolayer of biliary epithelial cells, in a 2D differentiation condition, but also form functional ductal structures in a 3D condition. Importantly, this EpCAM high spheroid based biliary tissue generation is significantly faster than other existing methods and does not require cell sorting. In addition, we show that a knock-in CK7 reporter human iPSC line generated by CRIPSR/Cas9 genome editing technology greatly facilitates the analysis of biliary differentiation. This new ductal differentiation method will provide a more efficient method of obtaining bile duct cells and tissues, which may facilitate engineering of complete and functional liver tissue in the future.
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影响因子：
13.5
作者：
Dianat, Noushin;Dubois-Pot-Schneider, Helene;Steichen, Clara;Desterke, Christophe;Leclerc, Philippe;Raveux, Aurelien;Combettes, Laurent;Weber, Anne;Corlu, Anne;Dubart-Kupperschmitt, Anne
通讯作者：
Dubart-Kupperschmitt, Anne
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
17.1
作者：
Liu H;Kim Y;Sharkis S;Marchionni L;Jang YY
通讯作者：
Jang YY
影响因子：
3.3
作者：
Tanimizu, Naoki;Miyajima, Atsushi;Mostov, Keith E.
通讯作者：
Mostov, Keith E.
影响因子：
46.9
作者：
Ogawa, Mina;Ogawa, Shinichiro;Ghanekar, Anand
通讯作者：
Ghanekar, Anand