Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates
Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates
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DOI：
10.3791/55403
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发表时间：
2017-03-01
影响因子：
1.2
通讯作者：
Hajduch, Marian
中科院分区：
文献类型：
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作者：
Das, Viswanath;Furst, Tomas;Hajduch, Marian
密切模仿体内条件的肿瘤模型在药物发现和开发中越来越受欢迎，以筛选潜在的抗癌药物。多细胞肿瘤球体 (MCTSes) 有效模拟实体瘤的生理条件，使其成为先导化合物优化和靶标验证的优秀体外模型。在可用于 MCTS 培养的各种技术中，琼脂糖上的液体覆盖方法是最便宜的 MCTS 生成方法之一。然而，使用液体覆盖进行高通量筛选的 MCTS 培养物的可靠转移可能会受到许多限制的影响，包括用琼脂糖包被微量滴定板 (MP) 以及跨孔形成均匀 MCTS 的不可重复性。 MP 很容易出现由于板外部介质过度蒸发而导致的边缘效应，从而妨碍使用整个板进行药物测试。本手稿对液体覆盖技术进行了详细的技术改进，以提高均匀 MCTS 形成的可扩展性和再现性。此外，还详细介绍了一种简单、半自动且普遍适用的软件工具，用于评估药物治疗后的 MCTS 特征。
Tumor models that closely imitate in vivo conditions are becoming increasingly popular in drug discovery and development for the screening of potential anti-cancer drugs. Multicellular tumor spheroids (MCTSes) effectively mimic the physiological conditions of solid tumors, making them excellent in vitro models for lead optimization and target validation. Out of the various techniques available for MCTS culture, the liquid-overlay method on agarose is one of the most inexpensive methods for MCTS generation. However, the reliable transfer of MCTS cultures using liquid-overlay for high-throughput screening may be compromised by a number of limitations, including the coating of microtiter plates (MPs) with agarose and the irreproducibility of uniform MCTS formation across wells. MPs are significantly prone to edge effects that result from excessive evaporation of medium from the exterior of the plate, preventing the use of the entire plate for drug tests. This manuscript provides detailed technical improvements to the liquid-overlay technique to increase the scalability and reproducibility of uniform MCTS formation. Additionally, details on a simple, semi-automatic, and universally applicable software tool for the evaluation of MCTS features after drug treatment is presented.