Refining In Vitro Toxicity Models: Comparing Baseline Characteristics of Lung Cell Types
Refining In Vitro Toxicity Models: Comparing Baseline Characteristics of Lung Cell Types
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DOI：
10.1093/toxsci/kfz001
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发表时间：
2019-04-01
影响因子：
3.8
通讯作者：
Sayes, Christie M.
中科院分区：
文献类型：
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作者：
Lujan, Henry;Criscitiello, Michael F.;Sayes, Christie M.
在体外毒理学领域，不断发展的需要是提高实验设计的质量，即从不明确的细胞培养到明确表征的细胞毒理学模型。随着环境健康科学家开始在肺毒理学研究中更加依赖细胞培养模型，这种演变尤为重要。这项研究分析了来自人类肺部两个不同深度的细胞的差异和相似性，这些细胞具有不同的表型。我们比较了上、下气道的原代、转化和癌源性细胞系的细胞周期和抗氧化剂相关mRNA和蛋白浓度。总之，本研究纳入了体外毒理学研究论文中报告的6种最常用的细胞系（即PTBE、BEAS-2B、A549、PSAE、Met-5A和Calu-3）。通过分子生物学（q-PCR、ELISA和流式细胞术）和显微镜（相位和荧光）技术以及细胞氧化应激终点分析完成细胞特征的比较。在使用统计分析比较每种细胞类型的反应后，结果证实了细胞周期调节剂的背景水平、固有抗氧化能力、促炎状态和不同毒理学反应的显著差异。分析的数据提高了我们对细胞特性的理解，反过来，有助于更准确地解释毒理学结果。我们的结论表明，在体外毒理学研究应包括一个详细的细胞表征部分发表的论文。
There is an ever-evolving need in the field of in vitro toxicology to improve the quality of experimental design, ie, from ill-defined cell cultures to well-characterized cytotoxicological models. This evolution is especially important as environmental health scientists begin to rely more heavily on cell culture models in pulmonary toxicology studies. The research presented in this study analyzes the differences and similarities of cells derived from two different depths of the human lung with varying phenotypes. We compared cell cycle and antioxidant-related mRNA and protein concentrations of primary, transformed, and cancer-derived cell lines from the upper and lower airways. In all, six of the most commonly used cell lines reported in in vitro toxicology research papers were included in this study (ie, PTBE, BEAS-2B, A549, PSAE, Met-5A, and Calu-3). Comparison of cell characteristics was accomplished through molecular biology (q-PCR, ELISA, and flow cytometry) and microscopy (phase and fluorescence) techniques as well as cellular oxidative stress endpoint analyses. After comparing the responses of each cell type using statistical analyses, results confirmed significant differences in background levels of cell cycle regulators, inherent antioxidant capacity, pro-inflammatory status, and differential toxicological responses. The analyzed data improve our understanding of the cell characteristics, and in turn, aids in more accurate interpretation of toxicological results. Our conclusions suggest that in vitro toxicology studies should include a detailed cell characterization component in published papers.