Scientific Product Assessment Center, R&D Group, Japan Tobacco Inc., 6-2 Umegaoka, Aoba-ku, Yokohama, Kanagawa 227-8512, Japan.
Scientific and Regulatory Affairs, JT International SA, 8 rue Kazem Radjavi, 1202 Geneva, Switzerland.
Toxicol In Vitro. 2022 Aug;82:105391. doi: 10.1016/j.tiv.2022.105391. Epub 2022 May 18.
Three-dimensional (3D) cultured primary cells are used to predict the toxicity of substances towards humans because these 3D cultures closely mimic the physiological architecture of tissues. Nonetheless, it is important to consider primary-cell-specific variability for endpoint selection and appropriate evaluation of toxicity because donor-dependent characteristics may be retained even in in vitro cell cultures. In this report, 3D differentiated bronchial epithelial cells from three donors were used to investigate donor-to-donor variability, with an aqueous extract of cigarette smoke (CS) used as the test substance. Ciliary function, cytokine secretion, and histopathology, which are affected by CS, were examined, and transcriptomic analysis was also performed. The results revealed that interleukin-8 secretion and oxidative stress-related gene expression were consistently altered for all donors; however, their amplitudes varied. Moreover, one of the donors showed unique responses to CS, suggesting that this donor was an outlier. This donor showed intrinsic differences in histology, cytokine secretion, and gene expression profile. Such donors may help evaluate potential toxicological concerns and aid our understanding of disease pathogenesis. Conversely, these donors may confound toxicological assessment and endpoint selection. Fit-for-purpose handling of inter-donor variability is warranted.
三维（3D）培养的原代细胞被用于预测物质对人类的毒性，因为这些 3D 培养物非常接近组织的生理结构。尽管如此，考虑到原代细胞的特异性变异性对于终点选择和毒性的适当评估仍然很重要，因为即使在体外细胞培养中，供体依赖性特征也可能保留下来。在本报告中，使用来自三个供体的 3D 分化支气管上皮细胞来研究供体间的变异性，使用香烟烟雾（CS）的水提取物作为测试物质。检查了受 CS 影响的纤毛功能、细胞因子分泌和组织病理学，同时还进行了转录组分析。结果表明，所有供体的白细胞介素-8 分泌和与氧化应激相关的基因表达均发生持续改变；然而，它们的幅度不同。此外，其中一个供体对 CS 表现出独特的反应，表明该供体是一个异常值。该供体在组织学、细胞因子分泌和基因表达谱方面表现出内在差异。这些供体可能有助于评估潜在的毒理学问题，并帮助我们了解疾病的发病机制。相反，这些供体可能会混淆毒理学评估和终点选择。需要对供体间的变异性进行适合目的的处理。
