{"title":"High-Throughput Assessment of Mechanistic Toxicity of Chemicals in Miniaturized 3D Cell Culture","authors":"Pranav Joshi, Soo-Yeon Kang, Akshata Datar, Moo-Yeal Lee","doi":"10.1002/cptx.66","DOIUrl":null,"url":null,"abstract":"<p>High-content imaging (HCI) assays on two-dimensional (2D) cell cultures often do not represent <i>in vivo</i> characteristics accurately, thus reducing the predictability of drug toxicity/efficacy <i>in vivo</i>. On the other hand, conventional 3D cell cultures are relatively low throughput and possess difficulty in cell imaging. To address these limitations, a miniaturized 3D cell culture has been developed on a micropillar/microwell chip platform with human cells encapsulated in biomimetic hydrogels. Model compounds are used to validate human cell microarrays for high-throughput assessment of mechanistic toxicity. Main mechanisms of toxicity of compounds can be investigated by analyzing multiple parameters such as DNA damage, mitochondrial impairment, intracellular glutathione level, and cell membrane integrity. IC<sub>50</sub> values of these parameters can be determined and compared for the compounds to investigate the main mechanism of toxicity. This paper describes miniaturized HCI assays on 3D-cultured cell microarrays for high-throughput assessment of mechanistic profiles of compound-induced toxicity. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"79 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.66","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in toxicology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cptx.66","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 2
Abstract
High-content imaging (HCI) assays on two-dimensional (2D) cell cultures often do not represent in vivo characteristics accurately, thus reducing the predictability of drug toxicity/efficacy in vivo. On the other hand, conventional 3D cell cultures are relatively low throughput and possess difficulty in cell imaging. To address these limitations, a miniaturized 3D cell culture has been developed on a micropillar/microwell chip platform with human cells encapsulated in biomimetic hydrogels. Model compounds are used to validate human cell microarrays for high-throughput assessment of mechanistic toxicity. Main mechanisms of toxicity of compounds can be investigated by analyzing multiple parameters such as DNA damage, mitochondrial impairment, intracellular glutathione level, and cell membrane integrity. IC50 values of these parameters can be determined and compared for the compounds to investigate the main mechanism of toxicity. This paper describes miniaturized HCI assays on 3D-cultured cell microarrays for high-throughput assessment of mechanistic profiles of compound-induced toxicity. © 2018 by John Wiley & Sons, Inc.
微型三维细胞培养中化学物质机械毒性的高通量评估
二维(2D)细胞培养物的高含量成像(HCI)检测通常不能准确地代表体内特征,从而降低了药物在体内毒性/疗效的可预测性。另一方面,传统的三维细胞培养通量相对较低,并且在细胞成像方面存在困难。为了解决这些限制,在微柱/微孔芯片平台上开发了一种小型化的3D细胞培养物,将人体细胞封装在仿生水凝胶中。模型化合物用于验证人体细胞微阵列的高通量评估机械毒性。通过分析DNA损伤、线粒体损伤、细胞内谷胱甘肽水平和细胞膜完整性等参数,可以探讨化合物毒性的主要机制。这些参数的IC50值可以用来测定和比较化合物的主要毒性机制。本文描述了在3d培养细胞微阵列上的小型化HCI实验,用于高通量评估化合物诱导毒性的机制概况。©2018 by John Wiley &儿子,Inc。
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