Yamashita Tomoki, Takayama Kazuo, Sakurai Fuminori, Mizuguchi Hiroyuki
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan; PRESTO, Japan Science and Technology Agency, Saitama 332-0012, Japan; Laboratory of Hepatocyte Regulation, National Institute of Biomedical Innovation, Health and Nutrition, Osaka 567-0085, Japan.
Biochem Biophys Res Commun. 2018 Feb 19;496(4):1269-1275. doi: 10.1016/j.bbrc.2018.01.186. Epub 2018 Feb 1.
Human induced pluripotent stem (iPS) cell-derived hepatocyte-like cells are expected to be utilized in drug screening and regenerative medicine. However, hepatocyte-like cells have not been fully used in such applications because it is difficult to produce such cells on a large scale. In this study, we tried to establish a method to mass produce hepatocyte-like cells using a three-dimensional (3D) cell culture bioreactor called the Rotary Cell Culture System (RCCS). RCCS enabled us to obtain homogenous hepatocyte-like cells on a billion scale (>10 cells). The gene expression levels of some hepatocyte markers (alpha-1 antitrypsin, cytochrome (CYP) 1A2, CYP2D6, and hepatocyte nuclear factor 4alpha) were higher in 3D-cultured hepatocyte-like cells than in 2D-cultured hepatocyte-like cells. This result suggests that RCCS could provide more suitable conditions for hepatocyte maturation than the conventional 2D cell culture conditions. In addition, more than 90% of hepatocyte-like cells were positive for albumin and could uptake low-density lipoprotein in the culture medium. We succeeded in the large-scale production of homogenous and functional hepatocyte-like cells from human iPS cells. This technology will be useful in drug screening and regenerative medicine, which require enormous numbers of hepatocyte-like cells.
人诱导多能干细胞（iPS）来源的肝样细胞有望用于药物筛选和再生医学。然而，肝样细胞尚未在这类应用中得到充分利用，因为大规模生产此类细胞存在困难。在本研究中，我们尝试建立一种利用名为旋转细胞培养系统（RCCS）的三维（3D）细胞培养生物反应器大规模生产肝样细胞的方法。RCCS使我们能够在十亿规模（>10⁹个细胞）上获得均匀的肝样细胞。一些肝细胞标志物（α-1抗胰蛋白酶、细胞色素（CYP）1A2、CYP2D6和肝细胞核因子4α）在3D培养的肝样细胞中的基因表达水平高于2D培养的肝样细胞。这一结果表明，与传统的2D细胞培养条件相比，RCCS可为肝细胞成熟提供更适宜的条件。此外，超过90%的肝样细胞白蛋白呈阳性，并且能够摄取培养基中的低密度脂蛋白。我们成功地从人iPS细胞大规模生产出了均匀且具有功能的肝样细胞。这项技术将有助于药物筛选和再生医学，因为这些领域需要大量的肝样细胞。
