Perfusion of medium with supplemented growth factors changes metabolic activities and cell morphology of hepatocyte-nonparenchymal cell coculture
Perfusion of medium with supplemented growth factors changes metabolic activities and cell morphology of hepatocyte-nonparenchymal cell coculture
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DOI：
10.1089/ten.2004.10.1297
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发表时间：
2004-09-01
影响因子：
--
通讯作者：
Ohshima, N
中科院分区：
文献类型：
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作者：
Kan, P;Miyoshi, H;Ohshima, N
为研制一种可行的灌注型生物人工肝装置，采用肝细胞生长因子（HGF）和肝素结合表皮生长因子样生长因子（HB-EGF）对肝细胞-非实质细胞（NPC）共培养物进行灌注。在培养的第1天，在1.3 dyn/cm（2）的恒定剪切应力下灌注增强了肝细胞的氨代谢和尿素合成活性。这些增强的活动持续到第7天，只有当生长因子存在。相反，在静态培养中没有观察到生长因子对这些活动的有益影响。在灌注培养中，形成三维细胞聚集体。在这些聚集体的表面上，发现主要由NPC组成的扁平细胞层，并且细胞聚集体的中心簇由圆形肝细胞和网硬蛋白原纤维组成。这些观察结果有力地表明，不同类型的肝细胞和结缔组织的重建在灌注培养中形成了能够调节肝细胞的肝脏特异性功能的仿组织细胞聚集体。因此，肝细胞- NPC共培养系统的灌注培养条件应适当设计，以诱导培养细胞的适当重建，以用作生物人工肝装置。
To develop a feasible perfusion-type bioartificial liver device, perfusion of hepatocyte - nonparenchymal cell (NPC) cocultures with medium supplemented with hepatocyte growth factor (HGF) and heparin-binding epidermal growth factor-like growth factor (HB-EGF) was carried out. On day 1 of culture, perfusion at a constant shear stress of 1.3 dyn/cm(2) enhanced ammonia metabolic and urea synthetic activities of hepatocytes. These enhanced activities were sustained up to day 7 only when growth factors were present. In contrast, no beneficial effects of growth factors on these activities were observed in static cultures. In perfusion cultures, three-dimensional cell aggregates were formed. On the surface of these aggregates, flattened cell layers composed mainly of NPCs were found, and the central cluster of cell aggregates was composed of round-shaped hepatocytes and reticulin fibrils. These observations strongly suggested that the reconstruction of different types of liver cells and connective tissues formed tissue-mimicking cell aggregates in the perfusion culture that was able to modulate the liver-specific functions of hepatocytes. Thus, perfusion culture conditions of the hepatocyte - NPC coculture system should be appropriately designed to induce suitable reconstruction of the cultured cells for use as a bioartificial liver device.