Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells.
Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells.
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DOI：
10.1038/nprot.2017.011
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发表时间：
2017-04
期刊：
影响因子：
14.8
通讯作者：
Vallier L
中科院分区：
文献类型：
--
作者：
Sampaziotis F;de Brito MC;Geti I;Bertero A;Hannan NR;Vallier L
分离和繁殖功能原代胆管细胞的困难是研究胆道疾病和测试新的治疗药物的主要限制。为了克服这一问题，我们开发了一个将人类多能干细胞（hPSCs）分化为功能性胆管细胞样细胞（CLCs）的平台。我们之前报道过，我们的26天方案紧密概括了胆道发育的关键阶段，从造血干细胞分化为内胚层和随后的前肠祖细胞开始，接着是肝母细胞的产生，表达早期胆道标志物的胆管细胞祖细胞和显示胆管细胞功能的成熟clc。与其他的hpsc胆道分化方案相比，我们的系统不需要与其他细胞类型共培养，并且依赖于化学定义的条件，直至并包括胆管细胞祖细胞的产生。复杂的细胞外基质用于clc的成熟，因此hPSC培养和3D类器官系统的经验可能是获得最佳结果的必要条件。最后，我们的平台产生大量疾病特异性功能胆管细胞的能力将在胆管病变，疾病建模和治疗化合物筛选方面具有广泛的应用。
The difficulty in isolating and propagating functional primary cholangiocytes is a major limitation in studying biliary disorders and testing novel therapeutic agents. To overcome this problem, we have developed a platform for the differentiation of human Pluripotent Stem Cells (hPSCs) into functional cholangiocyte-like cells (CLCs). We have previously reported that our 26-day protocol closely recapitulates key stages of biliary development starting with the differentiation of hPSCs into endoderm and subsequently foregut progenitor cells, followed by the generation of hepatoblasts, cholangiocyte progenitors expressing early biliary markers and mature CLCs displaying cholangiocyte functionality. Compared to alternative protocols for biliary differentiation of hPSCs, our system does not require co-culture with other cell types and relies on chemically defined conditions up to and including the generation of cholangiocyte progenitors. A complex extracellular matrix is used for the maturation of CLCs, therefore experience in hPSC culture and 3D organoid systems may be necessary for optimal results. Finally, the capacity of our platform for generating large amounts of disease-specific functional cholangiocytes will have broad applications for cholangiopathies, in disease modeling and for screening of therapeutic compounds.