The bioartificial liver (BAL) device is an extracorporeal liver support system incorporating living hepatocytes. A major problem in BAL device development is to obtain a high number of functional cells. In this study, we focused on a genetically engineered mouse hepatoma cell line, Hepa/8F5, in which elevated liver functions are induced via overexpression of liver-enriched transcription factors activated by doxycycline (Dox) addition. We applied a three-dimensional culture technique using hollow fibers (HFs) to Hepa/8F5 cells. Hepa/8F5 cells responded to Dox addition by reducing their proliferative activity and performing liver-specific functions of ammonia removal and albumin secretion. The functional activities of cells depended on the timing of Dox addition. We also found that Hepa/8F5 cells in the HF culture were highly functional in a low rather than high cell density environment. We further fabricated an HF-type bioreactor with immobilized Hepa/8F5 cells as a BAL device. Although ammonia removal activity of this BAL device was lower than that of the small-scale HF bundle, albumin secretion activity was slightly higher. These results indicated that the BAL device with immobilized Hepa/8F5 cells was highly functional with potential to show curative effects in liver failure treatment.

中文翻译：

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一种基于中空纤维的基因工程肝癌细胞三维培养的生物人工肝装置。

生物人工肝（BAL）设备是结合了活肝细胞的体外肝支持系统。BAL设备开发中的主要问题是获得大量功能单元。在这项研究中，我们专注于基因工程小鼠肝癌细胞系Hepa / 8F5，其中通过强力霉素（Dox）添加激活的肝脏富集转录因子的过表达诱导肝功能升高。我们将使用空心纤维（HF）的三维培养技术应用于Hepa / 8F5细胞。Hepa / 8F5细胞通过降低其增殖活性并执行除氨和清蛋白分泌的肝脏特异性功能来对Dox的添加做出反应。细胞的功能活性取决于添加Dox的时间。我们还发现，HF培养中的Hepa / 8F5细胞在低而不是高细胞密度的环境中具有很高的功能。我们进一步制造了固定化的Hepa / 8F5细胞作为BAL装置的HF型生物反应器。尽管该BAL装置的除氨活性低于小规模的HF束，但白蛋白分泌活性稍高。这些结果表明，具有固定化的Hepa / 8F5细胞的BAL装置具有很高的功能，并有望在肝衰竭治疗中显示出疗效。