Laboratory of Stem Cells and Translational Medicine, Institutes for Life Sciences, School of Medicine, South China University of Technology, Guangzhou 510006, China.
School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou 510180, China.
Cells. 2022 Dec 18;11(24):4117. doi: 10.3390/cells11244117.
Hepatocytes exhibit a multi-polarized state under the in vivo physiological environment, however, human embryonic stem cell-derived hepatocytes (hEHs) rarely exhibit polarity features in a two-dimensional (2D) condition. Thus, we hypothesized whether the polarized differentiation might enhance the maturity and liver function of hEHs. In this study, we obtained the polarized hEHs (phEHs) by using 2D differentiation in conjunct with employing transwell-based polarized culture. Our results showed that phEHs directionally secreted albumin, urea and bile acids, and afterward, the apical membrane and blood-bile barrier (BBIB) were identified to form in phEHs. Moreover, phEHs exhibited a higher maturity and capacitity of cellular secretory and drug metabolism than those of non-phEHs. Through transcriptome analysis, it was found that the polarized differentiation induced obvious changes in gene expression profiles of cellular adhesion and membrane transport in hEHs. Our further investigation revealed that the activation of Hippo and AMPK signaling pathways made contributions to the regulation of function and cellular polarity in phEHs, further verifying that the liver function of hEHs was closely related with their polarization state. These results not only demonstrated that the polarized differentiation enhanced the maturity and liver function of hEHs, but also identified the molecular targets that regulated the polarization state of hEHs.
在体内生理环境下，肝细胞呈现出多极状态，然而，人胚胎干细胞来源的肝细胞（hEHs）在二维（2D）条件下很少表现出极性特征。因此，我们假设极性分化是否可以增强 hEHs 的成熟度和肝功能。在这项研究中，我们通过使用 2D 分化并结合基于 Transwell 的极化培养来获得极化的 hEHs（phEHs）。我们的结果表明，phEHs 定向分泌白蛋白、尿素和胆汁酸，随后在 phEHs 中鉴定出顶膜和血胆汁屏障（BBIB）的形成。此外，phEHs 表现出比非 phEHs 更高的成熟度和细胞分泌以及药物代谢能力。通过转录组分析，发现极化分化诱导 hEHs 中细胞黏附和膜转运相关基因表达谱发生明显变化。我们进一步的研究表明，Hippo 和 AMPK 信号通路的激活有助于调节 phEHs 的功能和细胞极性，进一步证实 hEHs 的肝功能与其极化状态密切相关。这些结果不仅表明极化分化增强了 hEHs 的成熟度和肝功能，而且还确定了调节 hEHs 极化状态的分子靶标。
