Human stem cell-derived hepatocyte-like cells (HLCs) offer an attractive platform to study liver biology. Despite their numerous advantages, HLCs lack critical in vivo characteristics, including cell polarity. Here, we report a stem cell differentiation protocol that uses transwell filters to generate columnar polarized HLCs with clearly defined basolateral and apical membranes separated by tight junctions. We show that polarized HLCs secrete cargo directionally: Albumin, urea, and lipoproteins are secreted basolaterally, whereas bile acids are secreted apically. Further, we show that enterically transmitted hepatitis E virus (HEV) progeny particles are secreted basolaterally as quasi-enveloped particles and apically as naked virions, recapitulating essential steps of the natural infectious cycle in vivo. We also provide proof-of-concept that polarized HLCs can be used for pharmacokinetic and drug-drug interaction studies. This novel system provides a powerful tool to study hepatocyte biology, disease mechanisms, genetic variation, and drug metabolism in a more physiologically relevant setting.

人类干细胞衍生的肝细胞样细胞（HLC）为研究肝脏生物学提供了一个有吸引力的平台。尽管 HLC 具有众多优点，但其缺乏关键的体内特征，包括细胞极性。在这里，我们报告了一种干细胞分化方案，该方案使用 Transwell 过滤器生成柱状偏振 HLC，其具有明确定义的基底外侧膜和顶膜，由紧密连接分隔开。我们发现极化的 HLC 定向分泌货物：白蛋白、尿素和脂蛋白在基底外侧分泌，而胆汁酸在顶部分泌。此外，我们还发现，肠道传播的戊型肝炎病毒（HEV）子代颗粒在基底外侧分泌为准包膜颗粒，在顶部分泌为裸露病毒颗粒，重现了体内自然感染周期的基本步骤。我们还提供了极化 HLC 可用于药代动力学和药物相互作用研究的概念验证。这个新颖的系统提供了一个强大的工具，可以在更生理相关的环境中研究肝细胞生物学、疾病机制、遗传变异和药物代谢。