Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD, USA.
Methods Mol Biol. 2024;2749:85-90. doi: 10.1007/978-1-0716-3609-1_8.
Accurate evaluation of potential drug risks such as drug-induced liver injury (DILI) continues to be a challenge faced by pharmaceutical industry and regulatory agencies. Preclinical testing has served as a foundation for the evaluation of the potential risks and effectiveness of investigational new drug (IND) products in humans. However, current two-dimensional (2D) in vitro human primary hepatocyte (HPH) culture systems cannot accurately depict and simulate the rich environment and complex processes observed in vivo, while animal studies present inherited species-specific differences and low throughput scales. Thus, there is a continued demand to establish new approaches that can better characterize DILI during drug discovery and development. Among others, the three-dimensional (3D) hepatic spheroid model comprising self-aggregated primary human hepatocytes cocultured with non-parenchymal cells (NPCs) appears to be a more accurate representation of the natural hepatic microenvironment with intercellular interactions between hepatocytes, stellate cells, Kupffer cells, liver sinusoidal endothelial cells (LSECs), and other cell types. This model holds the potential to improve the ability for long-term functional and toxicological studies. Here, we provide methodological details for this human hepatic spheroid coculture model system.
准确评估药物引起的肝损伤（DILI）等潜在药物风险，一直是制药行业和监管机构面临的挑战。临床前测试是评估新研发药物（IND）产品在人体中潜在风险和疗效的基础。然而，目前的二维（2D）体外人原代肝细胞（HPH）培养系统无法准确描绘和模拟体内观察到的丰富环境和复杂过程，而动物研究则存在遗传的物种特异性差异和低通量规模。因此，人们一直需要建立新的方法，以便在药物发现和开发过程中更好地描述 DILI。其中，由自我聚集的原代人肝细胞与非实质细胞（NPC）共培养而成的三维（3D）肝球体模型，似乎更能准确地反映具有细胞间相互作用的天然肝微环境，包括肝细胞、星状细胞、库普弗细胞、肝窦内皮细胞（LSEC）和其他细胞类型。该模型有潜力提高长期功能和毒理学研究的能力。在这里，我们提供了该人肝球体共培养模型系统的方法学细节。
