Generation of functional cholangiocyte-like cells from human pluripotent stem cells and HepaRG cells.
Generation of functional cholangiocyte-like cells from human pluripotent stem cells and HepaRG cells.
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DOI：
10.1002/hep.27165
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发表时间：
2014-08
期刊：
影响因子：
13.5
通讯作者：
Dubart-Kupperschmitt, Anne
中科院分区：
文献类型：
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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
胆管细胞是胆管上皮细胞，与肝细胞一样，起源于胚胎发育过程中的肝母细胞。在这项研究中，我们研究了人胚胎干细胞(HESCs)向胆管细胞分化的可能性，并报道了一种新的方法，利用无饲养层和特定的培养条件推动hESCs向胆管细胞系分化。在分化为肝祖细胞后，用生长激素、表皮生长因子、白介素6和牛磺胆酸钠进一步分化为胆管细胞。这些条件也使我们能够从HepaRG来源的肝母细胞中产生胆管细胞。HESC和HepaRG来源的胆管细胞样细胞表达胆管细胞标志物，包括细胞角蛋白7和骨桥蛋白，转录因子SOX9和肝细胞核因子6。这些细胞还表达对胆管细胞功能重要的特异性蛋白，包括囊性纤维化跨膜传导调节因子、分泌素受体和核受体。它们形成初级纤毛，并通过增加细胞内钙离子对激素刺激做出反应。我们通过整合基因组学证明，标志hESC-或HepaRG-胆管细胞的基因的表达将肝细胞谱系与胆管细胞谱系分开。当在3D基质中生长时，胆管细胞形成上皮/基底膜两极，并形成功能性囊泡和胆管。此外，我们还展示了人类诱导的多能干细胞也可以生成胆管细胞样细胞，这证明了我们的方法对不同来源的干/祖细胞的有效性。结论：我们建立了一种强大而有效的方法将多能干细胞分化为胆管细胞样细胞，其结构和功能与正常肝脏的胆管细胞相似。这些细胞将有助于体外研究胆管发育的分子机制，并具有重要的治疗策略，包括生物工程肝脏方法。(《肝病》2014；60：700-714)
Cholangiocytes are biliary epithelial cells, which, like hepatocytes, originate from hepatoblasts during embryonic development. In this study we investigated the potential of human embryonic stem cells (hESCs) to differentiate into cholangiocytes and we report a new approach, which drives differentiation of hESCs toward the cholangiocytic lineage using feeder-free and defined culture conditions. After differentiation into hepatic progenitors, hESCs were differentiated further into cholangiocytes using growth hormone, epidermal growth factor, interleukin-6, and then sodium taurocholate. These conditions also allowed us to generate cholangiocytes from HepaRG-derived hepatoblasts. hESC- and HepaRG-derived cholangiocyte-like cells expressed markers of cholangiocytes including cytokeratin 7 and osteopontin, and the transcription factors SOX9 and hepatocyte nuclear factor 6. The cells also displayed specific proteins important for cholangiocyte functions including cystic fibrosis transmembrane conductance regulator, secretin receptor, and nuclear receptors. They formed primary cilia and also responded to hormonal stimulation by increase of intracellular Ca2+. We demonstrated by integrative genomics that the expression of genes, which signed hESC- or HepaRG-cholangiocytes, separates hepatocytic lineage from cholangiocyte lineage. When grown in a 3D matrix, cholangiocytes developed epithelial/apicobasal polarity and formed functional cysts and biliary ducts. In addition, we showed that cholangiocyte-like cells could also be generated from human induced pluripotent stem cells, demonstrating the efficacy of our approach with stem/progenitor cells of diverse origins. Conclusion: We have developed a robust and efficient method for differentiating pluripotent stem cells into cholangiocyte-like cells, which display structural and functional similarities to bile duct cells in normal liver. These cells will be useful for the in vitro study of the molecular mechanisms of bile duct development and have important potential for therapeutic strategies, including bioengineered liver approaches. (Hepatology 2014;60:700–714)
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影响因子：
17.1
作者：
Andersen JB;Factor VM;Marquardt JU;Raggi C;Lee YH;Seo D;Conner EA;Thorgeirsson SS
通讯作者：
Thorgeirsson SS
影响因子：
7.3
作者：
Ros, JE;Libbrecht, L;Roskams, TAD
通讯作者：
Roskams, TAD
影响因子：
13.5
作者：
Cerec, Virginie;Glaise, Denise;Corlu, Anne
通讯作者：
Corlu, Anne
影响因子：
24.5
作者：
Reynaert, H;Rombouts, K;Geerts, A
通讯作者：
Geerts, A
DOI：
10.1007/3-540-27194-5_2
发表时间：
2005-01-01
期刊：
SIGNALING PATHWAYS IN LIVER DISEASES
影响因子：
--
作者：
Leite, MF;Nathanson, MH
通讯作者：
Nathanson, MH