Liver progenitor cells develop cholangiocyte-type epithelial polarity in three-dimensional culture
Liver progenitor cells develop cholangiocyte-type epithelial polarity in three-dimensional culture
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DOI：
10.1091/mbc.e06-09-0848
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发表时间：
2007-04-01
影响因子：
3.3
通讯作者：
Mostov, Keith E.
中科院分区：
文献类型：
--
作者：
Tanimizu, Naoki;Miyajima, Atsushi;Mostov, Keith E.
胆管细胞是肝脏胆管系统的细胞组分，其起源于胚胎肝脏发育期间的成肝细胞。虽然一些转录因子和信号分子与胆管发育有关，但其分子机制尚未得到详细研究。在这里，我们应用三维（3D）培养技术的肝祖细胞系，HPPL，建立一个体外培养系统，HPPL获得分化的胆管细胞的特性。当HPPL生长在含有基质胶的凝胶中时，其中含有基底膜的细胞外基质成分，HPPL发展为顶基底极性并形成囊，囊内具有腔空间。在囊肿中，F-肌动蛋白束和非典型蛋白激酶C位于顶膜，E-钙粘蛋白位于侧膜，β-连环蛋白和整合素α 6位于底侧膜。囊肿中的HPPL表达胆管细胞标记物，包括细胞角蛋白19、整合素β 4和水通道蛋白-1，但不表达肝细胞标记物白蛋白。此外，HPPL运输罗丹明123，多药耐药基因产物的底物，从基底侧到中央腔。这些数据表明HPPL在3D培养中形成胆管细胞型上皮极性。磷脂酰肌醇3-激酶信号是必不可少的增殖和生存的HPPL在文化中，而层粘连蛋白-1是一个重要组成部分的基质胶诱导上皮极化的HPPL。由于HPPL囊肿显示与胆管的结构和功能相似性，HPPL的三维培养重现了体内胆管细胞分化，有助于研究体外胆管发育的分子机制。
Cholangiocytes are cellular components of the bile duct system of the liver, which originate from hepatoblasts during embryonic liver development. Although several transcription factors and signaling molecules have been implicated in bile duct development, its molecular mechanism has not been studied in detail. Here, we applied a three-dimensional (3D) culture technique to a liver progenitor cell line, HPPL, to establish an in vitro culture system in which HPPL acquire differentiated cholangiocyte characteristics. When HPPL were grown in a gel containing Matrigel, which contains extracellular matrix components of basement membrane, HPPL developed apicobasal polarity and formed cysts, which had luminal space inside. In the cysts, F-actin bundles and atypical protein kinase C were at the apical membrane, E-cadherin was localized at the lateral membrane, and beta-catenin and integrin alpha 6 were located at the basolateral membrane. HPPL in cysts expressed cholangiocyte markers, including cytokeratin 19, integrin beta 4, and aquaporin-1, but not a hepatocyte marker, albumin. Furthermore, HPPL transported rhodamine 123, a substrate for multidrug resistance gene products, from the basal side to the central lumen. These data indicate that HPPL develop cholangiocyte-type epithelial polarity in 3D culture. Phosphatidylinositol 3-kinase signaling was essential for proliferation and survival of HPPL in culture, whereas laminin-1 was a crucial component of Matrigel for inducing epithelial polarization of HPPL. Because HPPL cysts display structural and functional similarities with bile ducts, the 3D culture of HPPL recapitulates in vivo cholangiocyte differentiation and is useful to study the molecular mechanism of bile duct development in vitro.