Spontaneous hepatocyte migration towards an endothelial cell tube network
Spontaneous hepatocyte migration towards an endothelial cell tube network
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DOI：
10.1002/term.2577
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发表时间：
2018-03-01
影响因子：
3.3
通讯作者：
Eguchi, Susumu
中科院分区：
文献类型：
--
作者：
Baimakhanov, Zhassulan;Sakai, Yusuke;Eguchi, Susumu
工程化肝组织的一个关键部分是非实质细胞的贡献和维持复杂的三维（3D）结构在体外为他们的正常生理和功能。我们使用原代分离的大鼠肝细胞和来自人内皮静脉上皮细胞（HUVECs）的内皮细胞管网络生成3D肝组织。为了产生3D肝组织，在Matrigel（R）上进行原代肝细胞和管状结构HUVEC的共培养。在HUVEC形成管状结构后，将原代分离的大鼠肝细胞接种到HUVEC管状结构层上并培养24小时。我们研究了细胞迁移，细胞相互作用，和分布的HUVEC管结构和肝细胞使用多细胞成像培养箱，共聚焦显微镜和电子显微镜分析。在培养期间，延时成像显示肝细胞在凝胶中自发迁移，并且在24小时培养期后，绝大多数肝细胞已经移动并粘附到HUVEC管结构的表面。共聚焦显微镜测定证实了肝细胞和HUVEC管结构之间的这种独特的3D细胞相互作用。肝细胞能够保持其球形形状，以及HUVEC（具有管状腔的管状形式）。我们推测，肝细胞和内皮细胞的共培养复制了它们正常生理的一部分，并可能有助于诱导体外迁移和复杂生物组织结构的生长。
A crucial part of the engineering liver tissue is contribution of nonparenchymal cells and maintenance of a complex three-dimensional (3D) structure in vitro for their normal physiology and function. We generated 3D hepatic tissue using primary isolated rat hepatocytes and an endothelial cell tube network from human endothelial vein epithelial cells (HUVECs). To create the 3D hepatic tissue, coculture of primary hepatocytes and tube-structured HUVECs was performed on a Matrigel (R). After the HUVECs formed the tube structures, primary isolated rat hepatocytes were inoculated onto the HUVEC tube-structured layer and cultured for 24hr. We investigated the cell migration, cellular interaction, and distributions of HUVEC tube structures and hepatocytes using multi cell-imaging incubator, confocal microscopy, and electron microscopy analyses. During the culture time, time-lapse imaging showed spontaneous migration of the hepatocytes in the gel, and after the 24-hr culture period, the vast majority of the hepatocytes had moved and adhered to the surface of the HUVEC tube structures. A confocal microscopy assay confirmed this unique 3D cellular interaction between hepatocytes and HUVEC tube structures. The hepatocytes were able to maintain their spherical shape, as well as HUVECs (tube-like form with tubular cavity). We speculate that coculturing of hepatocytes and endothelial cells replicates part of their normal physiology and may help induce migration in vitro and the growth of complex biological tissue structures.