用单一或多种细胞类型产生的肝类器官已用于研究肝纤维化的发展、毒性、发病机制和药物筛选。然而，类器官的产生受到从原代组织中分离的细胞或从各种干细胞分化而来的细胞的可用性的限制。为了确保类器官形成的细胞可用性，我们研究了是否可以使用基于细胞系的 Huh-7 肝细胞癌细胞、THP-1 单核细胞分化的巨噬细胞、LX-2 肝星状细胞 (HSC) 和原代肝脏来生成肝脏类器官正弦内皮细胞（LSEC）。在肝脏类器官中，与二维 (2D) 培养的 Huh-7/LSEC/THP-1/LX-2 细胞（不含 Matrigel）相比，肝细胞、LSEC、巨噬细胞和 HSC 相关基因表达有所增加。硫代乙酰胺 (TAA) 增加了肝类器官中 α-平滑肌肌动蛋白的表达，但在 2D 培养的细胞中没有增加，而在 TAA 处理的类器官中，肝脏和 LSEC 标记物的表达减少，巨噬细胞和 HSC 标记物的表达增加。 N-乙酰基-L-半胱氨酸或肿瘤坏死因子刺激基因 6 蛋白治疗可抑制 TAA 诱导的纤维化。结果表明，肝脏毒物可诱导由Huh-7/LSEC/巨噬细胞/LX-2细胞组成的肝脏类器官发生纤维化和炎症反应，从而导致肝脏类器官纤维化。我们建议基于细胞系的类器官可用于疾病建模和药物筛选，以改善肝纤维化治疗。

Liver organoids generated with single or multiple cell types have been used to investigate liver fibrosis development, toxicity, pathogenesis, and drug screening. However, organoid generation is limited by the availability of cells isolated from primary tissues or differentiated from various stem cells. To ensure cell availability for organoid formation, we investigated whether liver organoids could be generated with cell-line-based Huh-7 hepatocellular carcinoma cells, macrophages differentiated from THP-1 monocytes, and LX-2 hepatic stellate cells (HSCs) and primary liver sinusoidal endothelial cells (LSECs). In liver organoids, hepatocyte-, LSEC-, macrophage-, and HSC-related gene expression increased relative to that in two-dimensional (2D)-cultured Huh-7/LSEC/THP-1/LX-2 cells without Matrigel. Thioacetamide (TAA) increased α-smooth muscle actin expression in liver organoids but not in 2D-cultured cells, whereas in TAA-treated organoids, the expression of hepatic and LSEC markers decreased and that of macrophage and HSC markers increased. TAA-induced fibrosis was suppressed by treatment with N-acetyl-L-cysteine or tumor-necrosis-factor-stimulated gene 6 protein. The results showed that liver toxicants could induce fibrotic and inflammatory responses in liver organoids comprising Huh-7/LSEC/macrophages/LX-2 cells, resulting in fibrotic liver organoids. We propose that cell-line-based organoids can be used for disease modeling and drug screening to improve liver fibrosis treatment.