开发概括关键肝功能的体外模型对于准确评估药物毒性至关重要。尽管肝脏类器官可用于药物发现和毒理学，但它们受到以下限制：（i）缺乏异生素代谢酶的表达和活性，以及​​（ii）难以模拟非酒精性脂肪肝病（NAFLD，它影响这些酶的表达）在体外。在这里，我们从 HepaRG 细胞、原代人巨噬细胞和肝星状细胞衍生的 LX-2 细胞生成了三维多细胞型肝脏类器官（以下简称“HML 类器官”）。我们还通过用硬脂酸和油酸的混合物培养 HML 类器官 9 天，开发了 NAFLD 模型。暴露的类器官表现出典型的脂肪变性特征并表达纤维化标志物。我们随后使用 HML 和 NAFLD-HML 类器官来模拟药物引起的肝损伤。通过估计 IC50 和基准剂量，我们能够改进可能对脂肪肝有毒的药物的体外检测。因此，HML 和 NAFLD-HML 类器官表现出大部分肝脏功能，并且是 NAFLD 中药物代谢、药物毒性和药物不良事件的相关体外模型。版权所有 © 2023。由 Elsevier Ltd 出版。

The development of in vitro models that recapitulate critical liver functions is essential for accurate assessments of drug toxicity. Although liver organoids can be used for drug discovery and toxicology, they are limited by (i) the lack of expression and activity of xenobiotic-metabolizing enzymes, and (ii) the difficulty of mimicking non-alcoholic fatty liver disease (NAFLD, which influences the expression of these enzymes) in vitro. Here, we generated three-dimensional multi-cell-type liver organoids (hereafter "HML organoids") from HepaRG cells, primary human macrophages, and hepatic-stellate-cell-derived LX-2 cells. We also developed an NAFLD model by culturing HML organoids for 9 days with a mixture of stearic and oleic acids. The exposed organoids showed typical features of steatosis and expressed fibrosis markers. We subsequently used HML and NAFLD-HML organoids to model drug-induced liver injury. By estimating the IC50 and benchmark doses, we were able to improve the in vitro detection of drugs likely to be toxic in fatty livers. Thus, HML and NAFLD-HML organoids exhibited most of the liver's functions and are relevant in vitro models of drug metabolism, drug toxicity, and adverse drug event in NAFLD.Copyright © 2023. Published by Elsevier Ltd.