High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array.
High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array.
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DOI：
10.1039/c0an00609b
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发表时间：
2011-02-07
期刊：
影响因子：
--
通讯作者：
Takayama S
中科院分区：
文献类型：
--
作者：
Tung YC;Hsiao AY;Allen SG;Torisawa YS;Ho M;Takayama S
细胞作为三维（3D）聚集体的培养可以增强基础生物学研究以及治疗方法开发的体外测试。然而，这样的3D培养模型通常比二维（2D）培养更复杂、麻烦和昂贵。本文描述了一种384孔格式的悬挂滴培养板，它可以在获得的3D细胞结构上进行球体形成，培养和随后的药物测试，就像传统的2D培养一样，可以直接执行并适应现有的高通量筛选（HTS）仪器。利用这个平台，我们发现与传统的二维单层细胞相比，具有不同作用模式的药物在生理三维细胞球体中产生不同的反应。具体来说，抗癌药物5-氟尿嘧啶（5-FU）对2D培养物具有更高的抗增殖作用，而缺氧激活药物通常被称为替拉帕嗪（TPZ）对3D培养物更有效。多路3D挂滴培养和测试板提供了一种有效的方法来获得通常在2D平台中丢失的生物学见解。
Culture of cells as three-dimensional (3D) aggregates can enhance in vitro tests for basic biological research as well as for therapeutics development. Such 3D culture models, however, are often more complicated, cumbersome, and expensive than two-dimensional (2D) cultures. This paper describes a 384-well format hanging drop culture plate that makes spheroid formation, culture, and subsequent drug testing on the obtained 3D cellular constructs as straightforward to perform and adapt to existing high-throughput screening (HTS) instruments as conventional 2D cultures. Using this platform, we show that drugs with different modes of action produce distinct responses in the physiological 3D cell spheroids compared to conventional 2D cell monolayers. Specifically, the anticancer drug 5-fluorouracil (5-FU) has higher anti-proliferative effects on 2D cultures whereas the hypoxia activated drug commonly referred to as tirapazamine (TPZ) are more effective against 3D cultures. The multiplexed 3D hanging drop culture and testing plate provides an efficient way to obtain biological insights that are often lost in 2D platforms.
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