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Abstract：
Chemicals in the systemic circulation can undergo hepatic xenobiotic metabolism, generate metabolites, and exhibit altered toxicity compared with their parent compounds. This article describes a 2-chamber liver-organ coculture model in a higher-throughput 96-well format for the determination of toxicity on target tissues in the presence of physiologically relevant human liver metabolism. This 2-chamber system is a hydrogel formed within each well consisting of a central well (target tissue) and an outer ring-shaped trough (human liver tissue). The target tissue chamber can be configured to accommodate a three-dimensional (3D) spheroid-shaped microtissue, or a 2-dimensional (2D) cell monolayer. Culture medium and compounds freely diffuse between the 2 chambers. Human-differentiated HepaRG liver cells are used to form the 3D human liver microtissues, which displayed robust protein expression of liver biomarkers (albumin, asialoglycoprotein receptor, Phase I cytochrome P450 [CYP3A4] enzyme, multidrug resistance-associated protein 2 transporter, and glycogen), and exhibited Phase I/II enzyme activities over the course of 17 days. Histological and ultrastructural analyses confirmed that the HepaRG microtissues presented a differentiated hepatocyte phenotype, including abundant mitochondria, endoplasmic reticulum, and bile canaliculi. Liver microtissue zonation characteristics could be easily modulated by maturation in different media supplements. Furthermore, our proof-of-concept study demonstrated the efficacy of this coculture model in evaluating testosterone-mediated androgen receptor responses in the presence of human liver metabolism. This liver-organ coculture system provides a practical, higher-throughput testing platform for metabolism-dependent bioactivity assessment of drugs/chemicals to better recapitulate the biological effects and potential toxicity of human exposures.
摘要：
体循环中的化学物质可以进行肝脏异种生物代谢，产生代谢物，与它们的母体化合物相比，表现出改变的毒性。本文介绍了一种高通量96孔格式的2室肝器官共培养模型，用于在存在生理相关的人类肝脏代谢的情况下确定对靶组织的毒性。该2室系统是在由中心孔(目标组织)和外部环形槽(人肝脏组织)组成的每个孔中形成的水凝胶。目标组织腔室可以被配置为容纳三维(3D)球形微组织，或二维(2D)细胞单层。培养基和化合物在2个室之间自由扩散。人分化的HepaRG肝细胞用于形成3D人肝微组织，显示肝脏生物标志物(白蛋白，去唾液酸糖蛋白受体，第一阶段细胞色素P450[CYP3A4]酶，多药耐药相关蛋白2转运体，和糖原），并在17天的过程中表现出I/II期酶活性。组织学和超微结构分析证实，HepaRG微组织呈现分化的肝细胞表型，包括丰富的线粒体，内质网，和胆管。可以通过在不同培养基补充剂中成熟来轻松调节肝微组织分区特征。此外，我们的概念验证研究证明了这种共培养模型在评估人肝脏代谢存在时睾酮介导的雄激素受体反应方面的有效性.这种肝脏-器官共培养系统提供了一种实用的，用于药物/化学品代谢依赖性生物活性评估的高通量测试平台，以更好地概括人类暴露的生物效应和潜在毒性。
