Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Laboratory of Hepatocyte Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan.
PLoS One. 2020 Feb 27;15(2):e0229654. doi: 10.1371/journal.pone.0229654. eCollection 2020.
Human hepatocytes are essential materials in pharmaceutical researches. Not only primary human hepatocytes (PHH) but also human iPS cell-derived hepatocyte-like cells (human iPS-HLCs) are expected to be applied as materials for pharmaceutical researches. To date, several culture media have been developed for culturing human hepatocytes. However, there have been no reports comparing these media to determine which is most suitable for culturing human hepatocytes. In this study, we compared five commercial media (Hepatocyte Culture Medium (HCM), HepatoZYME-SFM, Cellartis Power Primary HEP Medium, DMEM/F12, and William's E Medium (WEM)) to determine which is most suitable for culturing PHH and human iPS-HLCs. In hepatic differentiation of human iPS cells (day 14-25 of differentiation), albumin (ALB) and urea secretion abilities and CYP2C9, CYP2C19, and CYP3A4 activities were the highest when using HCM or WEM. During maintenance of human iPS-HLCs, ALB and urea producing abilities and CYP2C9, CYP2C19, and CYP3A4 activities were the highest when using HCM. Importantly, we found that human iPS-HLCs cultured in HCM were maintained for 3 weeks or more without impairment of their hepatic functions. These results suggest that it is necessary to select an optimal medium for hepatic differentiation and maintenance of human iPS-HLCs. In the case of PHH culture, there was little difference in hepatic functions among the five media. However, the CYP2C9, CYP2C19, and CYP3A4 activities were the highest when using HCM and WEM. In conclusion, it is important to select the optimal medium for specific application when carrying out pharmaceutical researches using human hepatocytes.
人源肝细胞是药物研究中的重要材料。不仅原代人肝细胞（PHH），而且人诱导多能干细胞衍生的肝细胞样细胞（hiPS-HLC）也有望作为药物研究的材料。迄今为止，已经开发了几种用于培养人肝细胞的培养基。然而，目前还没有比较这些培养基以确定哪种最适合培养人肝细胞的报道。在本研究中，我们比较了五种商业培养基（肝细胞培养基（HCM）、HepatoZYME-SFM、Cellartis Power Primary HEP 培养基、DMEM/F12 和 William's E 培养基（WEM）），以确定哪种最适合培养 PHH 和 hiPS-HLC。在人 iPS 细胞的肝分化（分化的第 14-25 天）过程中，使用 HCM 或 WEM 时白蛋白（ALB）和尿素分泌能力以及 CYP2C9、CYP2C19 和 CYP3A4 活性最高。在 hiPS-HLC 的维持过程中，使用 HCM 时 ALB 和尿素产生能力以及 CYP2C9、CYP2C19 和 CYP3A4 活性最高。重要的是，我们发现用 HCM 培养的 hiPS-HLC 可以维持 3 周或更长时间而不会损害其肝功能。这些结果表明，有必要为 hiPS-HLC 的肝分化和维持选择最佳培养基。在 PHH 培养的情况下，五种培养基之间的肝功能差异不大。然而，使用 HCM 和 WEM 时 CYP2C9、CYP2C19 和 CYP3A4 活性最高。总之，在使用人肝细胞进行药物研究时，选择最佳的培养基对于特定的应用非常重要。
