Department of Chemical Engineering, Faculty of Engineering, Kyushu University, Fukuoka 819-0395, Japan.
Cells. 2022 Apr 1;11(7):1194. doi: 10.3390/cells11071194.
Functional human hepatocytes have been a pivotal tool in pharmacological studies such as those investigating drug metabolism and hepatotoxicity. However, primary human hepatocytes are difficult to obtain in large quantities and may cause ethical problems, necessitating the development of a new cell source to replace human primary hepatocytes. We previously developed genetically modified murine hepatoma cell lines with inducible enhanced liver functions, in which eight liver-enriched transcription factor (LETF) genes were introduced into hepatoma cells as inducible transgene expression cassettes. Here, we establish a human hepatoma cell line with heat-inducible liver functions using HepG2 cells. The genetically modified hepatoma cells, designated HepG2/8F_HS, actively proliferated under normal culture conditions and, therefore, can be easily prepared in large quantities. When the expression of LETFs was induced by heat treatment at 43 °C for 30 min, cells ceased proliferation and demonstrated enhanced liver functions. Furthermore, three-dimensional spheroid cultures of HepG2/8F_HS cells showed a further increase in liver functions upon heat treatment. Comprehensive transcriptome analysis using DNA microarrays revealed that HepG2/8F_HS cells had enhanced overall expression of many liver function-related genes following heat treatment. HepG2/8F_HS cells could be useful as a new cell source for pharmacological studies and for constructing bioartificial liver systems.
功能人肝细胞一直是药理学研究的重要工具，例如药物代谢和肝毒性研究。然而，由于难以大量获得人原代肝细胞，且可能引发伦理问题，因此需要开发新的细胞来源来替代人原代肝细胞。我们之前开发了具有诱导增强肝功能的基因修饰鼠肝癌细胞系，其中 8 个肝富集转录因子（LETF）基因作为诱导转基因表达盒被引入肝癌细胞。在这里，我们使用 HepG2 细胞建立了一种具有热诱导肝功能的人肝癌细胞系。该基因修饰的肝癌细胞被命名为 HepG2/8F_HS，在正常培养条件下积极增殖，因此可以很容易地大量制备。当通过 43°C 热处理 30 分钟诱导 LETFs 的表达时，细胞停止增殖并表现出增强的肝功能。此外，HepG2/8F_HS 细胞的三维球体培养物在热处理后肝脏功能进一步增强。使用 DNA 微阵列进行的综合转录组分析表明，HepG2/8F_HS 细胞在热处理后整体表达了许多与肝功能相关的基因。HepG2/8F_HS 细胞可用作药理学研究和构建生物人工肝系统的新细胞来源。
