Ham Stephanie Lemmo, Atefi Ehsan, Fyffe Darcy, Tavana Hossein
Department of Biomedical Engineering, The University of Akron.
Department of Biomedical Engineering, The University of Akron;
J Vis Exp. 2015 Apr 23(98):e52754. doi: 10.3791/52754.
Cancer cell spheroids present a relevant in vitro model of avascular tumors for anti-cancer drug testing applications. A detailed protocol for producing both mono-culture and co-culture spheroids in a high throughput 96-well plate format is described in this work. This approach utilizes an aqueous two-phase system to confine cells into a drop of the denser aqueous phase immersed within the second aqueous phase. The drop rests on the well surface and keeps cells in close proximity to form a single spheroid. This technology has been adapted to a robotic liquid handler to produce size-controlled spheroids and expedite the process of spheroid production for compound screening applications. Spheroids treated with a clinically-used drug show reduced cell viability with increase in the drug dose. The use of a standard micro-well plate for spheroid generation makes it straightforward to analyze viability of cancer cells of drug-treated spheroids with a micro-plate reader. This technology is straightforward to implement both robotically and with other liquid handling tools such as manual pipettes.
癌细胞球体为抗癌药物测试应用提供了一种相关的无血管肿瘤体外模型。本文描述了一种在高通量96孔板格式中生产单培养和共培养球体的详细方案。该方法利用水两相系统将细胞限制在浸没于第二水相中的一滴较密水相中。液滴位于孔表面，使细胞紧密靠近以形成单个球体。该技术已被应用于机器人液体处理仪，以生产尺寸可控的球体，并加快用于化合物筛选应用的球体生产过程。用临床使用的药物处理的球体随着药物剂量的增加显示出细胞活力降低。使用标准微孔板生成球体使得用微孔板读数器分析药物处理的球体中癌细胞的活力变得简单直接。该技术无论是通过机器人操作还是与其他液体处理工具（如手动移液器）一起实施都很简单直接。
