A spheroid toxicity assay using magnetic 3D bioprinting and real-time mobile device-based imaging.
A spheroid toxicity assay using magnetic 3D bioprinting and real-time mobile device-based imaging.
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DOI：
10.1038/srep13987
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发表时间：
2015-09-14
影响因子：
4.6
通讯作者：
Souza GR
中科院分区：
文献类型：
--
作者：
Tseng H;Gage JA;Shen T;Haisler WL;Neeley SK;Shiao S;Chen J;Desai PK;Liao A;Hebel C;Raphael RM;Becker JL;Souza GR
生物医学研究中持续存在的挑战是寻找使用 3D 细胞培养物进行毒性筛查的简单而可靠的检测方法。这项研究通过一种新颖的球体测定来解决这一挑战，其中通过磁性 3D 生物打印形成的球体在细胞重新排列并压缩与活力和细胞骨架组织相关的球体时立即收缩。因此，球体大小可以用作毒性的简单度量。本研究的目的是验证球体收缩作为细胞毒性终点，使用 3T3 成纤维细胞响应 5 种有毒化合物（全反式视黄酸、地塞米松、多柔比星、5′-氟尿嘧啶、毛喉素）、十二烷基硫酸钠（+对照）和青霉素-G（-对照）。使用移动设备进行实时成像以提高吞吐量和效率。除青霉素-G 外，所有化合物均以剂量依赖性方式显着减缓收缩 (Z’ = 0.88)。 3D 细胞比 2D 细胞更能抵抗毒性，2D 细胞的毒性是通过 MTT 测定来测量的。球体的荧光染色和基因表达谱证实了这些发现。本研究的结果验证了该测定中的球体收缩是在代表性 3D 环境中进行高通量化合物筛选的简单且生物学相关的终点。
An ongoing challenge in biomedical research is the search for simple, yet robust assays using 3D cell cultures for toxicity screening. This study addresses that challenge with a novel spheroid assay, wherein spheroids, formed by magnetic 3D bioprinting, contract immediately as cells rearrange and compact the spheroid in relation to viability and cytoskeletal organization. Thus, spheroid size can be used as a simple metric for toxicity. The goal of this study was to validate spheroid contraction as a cytotoxic endpoint using 3T3 fibroblasts in response to 5 toxic compounds (all-trans retinoic acid, dexamethasone, doxorubicin, 5′-fluorouracil, forskolin), sodium dodecyl sulfate (+control), and penicillin-G (−control). Real-time imaging was performed with a mobile device to increase throughput and efficiency. All compounds but penicillin-G significantly slowed contraction in a dose-dependent manner (Z’ = 0.88). Cells in 3D were more resistant to toxicity than cells in 2D, whose toxicity was measured by the MTT assay. Fluorescent staining and gene expression profiling of spheroids confirmed these findings. The results of this study validate spheroid contraction within this assay as an easy, biologically relevant endpoint for high-throughput compound screening in representative 3D environments.