Robotic printing and drug testing of 384-well tumor spheroids.
Robotic printing and drug testing of 384-well tumor spheroids.
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384 孔肿瘤球体的机器人打印和药物测试。
DOI：
10.1109/embc.2015.7318823
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Tavana,Hossein
中科院分区：
文献类型：
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作者：
Ham,StephanieL;Thakuri,PradipS;Tavana,Hossein
抗癌药物开发的一个主要障碍是缺乏可靠和廉价的肿瘤模型来测试候选化合物的疗效。这一需求是由于广泛使用的单层培养不足以预测体内药物疗效而产生的。球体是一种3D致密的癌细胞簇，它模拟了肿瘤的重要特征，并为药物测试提供了组织模拟。在这里，我们提出了一种新的球体形成微技术，该技术简单易用，并允许在384微孔板中进行高通量药物筛选。这种方法是基于聚合物双水相体系。密度较高的水相以所需的密度与癌细胞混合。使用机器人液体处理机，将一滴这种细胞悬浮液分配到包含第二个浸泡水相的384微孔板的每一个孔中。癌细胞保持在位于井底的液滴中，并在孵化过程中形成一个椭圆形。液体输送机器人的使用确保了一滴水的精确分配，导致每口井只有一个椭球体，每个平板内的椭球体大小均匀。我们证实了球体产生的一致性，并证明了它们与肿瘤的生物学相关性。一项对标准化疗化合物阿霉素治疗的三阴性乳腺癌细胞球体的概念验证研究表明，这种方法在药物测试中具有潜力。与现有方法相比，这种球体培养微技术具有关键的优势，例如易于添加药物和活性试剂，能够在不将球体转移到新平板的情况下对其进行分析，以及不需要专门的平板或设备来形成球体。将这项技术纳入抗癌药物开发流程将有助于更有效地审查候选药物的疗效，并加快新药的发现。
A major impediment to anti-cancer drug development is the lack of a reliable and inexpensive tumor model to test the efficacy of candidate compounds. This need has emerged due to the insufficiency of widely-used monolayer cultures to predict drug efficacy in vivo. Spheroids, 3D compact clusters of cancer cells, mimic important characteristics of tumors and provide a tissue analog for drug testing. Here we present a novel spheroid formation microtechnology that is simple to use and allows high throughput drug screening in 384-microwell plates. This approach is based on a polymeric aqueous two-phase system. The denser aqueous phase is mixed with cancer cells at a desired density. Using a robotic liquid handler, a drop of this cell suspension is dispensed into each well of a 384-microwell plate containing the second, immersion aqueous phase. Cancer cells remain contained in the drop, which rests on the well bottom, and form a spheroid during incubation. The use of liquid handling robotics ensures precise dispensing of a single drop, resulting in a single spheroid per well and homogenously sized spheroids within each plate. We confirmed the consistency of production of spheroids and demonstrated their biological relevance to tumors. A proof of concept study with spheroids of triple negative breast cancer cells treated with a standard chemotherapeutic compound, doxorubicin, showed the potential of this method for drug testing. This spheroid culture microtechnology presents key advantages over existing methods such as the ease of drug and viability reagent addition, ability to analyze spheroids without transferring them to a new plate, and the elimination of the need for specialized plates or devices to form spheroids. Incorporating this technology in anti-cancer drug development pipeline will help examine the efficacy of drug candidates more effectively and expedite discovery of novel drugs.