Data from: From the cover: three-dimensional (3d) heparg spheroid model with physiologically relevant xenobiotic metabolism competence and hepatocyte functionality for liver toxicity screening.

Effective prediction of human responses to chemical and drug exposure is of critical importance in environmental toxicology research and drug development. While significant progress has been made to address this challenge using invitro liver models, these approaches often fail due to inadequate tissue model functionality. Herein, we describe the development, optimization, and characterization of a novel three-dimensional (3D) spheroid model using differentiated HepaRG cells that achieve and maintain physiologically relevant levels of xenobiotic metabolism (CYP1A2, CYP2B6, and CYP3A4/5). This invitro model maintains a stable phenotype over multiple weeks in both 96- and 384-well formats, supports highly reproducible tissue-like architectures and models pharmacologically- and environmentally important hepatic receptor pathways (ie AhR, CAR, and PXR) analogous to primary human hepatocyte cultures. HepaRG spheroid cultures use 50–100× fewer cells than conventional two dimensional cultures, and enable the identification of metabolically activated toxicants. Spheroid size, time in culture and culture media composition were important factors affecting basal levels of xenobiotic metabolism and liver enzyme inducibility with activators of hepatic receptors AhR, CAR and PXR. Repeated exposure studies showed higher sensitivity than traditional 2D cultures in identifying compounds that cause liver injury and metabolism-dependent toxicity. This platform combines the well-documented impact of 3D culture configuration for improved tissue functionality and longevity with the requisite throughput and repeatability needed for year-over-year toxicology screening.

精准预测人体对化学物与药物暴露的应答，在环境毒理学研究与药物开发领域具有至关重要的意义。尽管利用体外（in vitro）肝模型应对这一难题已取得显著进展，但此类方法常因组织模型功能不足而失效。本文详述了一种基于分化型HepaRG细胞的新型三维（3D）球体模型的开发、优化与表征，该模型可达到并维持符合生理学相关水平的外源性物质代谢活性（涵盖CYP1A2、CYP2B6及CYP3A4/5亚型）。该体外模型可在96孔与384孔板两种培养体系中维持数周的稳定表型，支持高度可复现的组织样结构构建，且可模拟与药理学及环境毒理学密切相关的肝受体通路（即芳烃受体（AhR）、组成型雄烷受体（CAR）及孕烷X受体（PXR）），其效果与原代人肝细胞培养体系类似。HepaRG球体培养体系所需细胞数仅为传统二维培养体系的1/50至1/100，且可用于识别经代谢活化的有毒物质。球体尺寸、培养时长及培养基组分是影响外源性物质基础代谢水平，以及肝受体AhR、CAR与PXR激活剂诱导肝酶活性的关键因素。重复暴露实验表明，相较于传统二维培养体系，该模型在鉴定可引发肝损伤及代谢依赖性毒性的化合物时具有更高的灵敏度。该平台整合了已被充分证实的三维培养体系优势——可提升组织功能与培养寿命，同时兼具高通量筛选与可重复性要求，可满足长期毒理学筛查的需求。