Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, China.
Anal Chem. 2013 Jul 16;85(14):6740-7. doi: 10.1021/ac400688f. Epub 2013 Jul 3.
We performed cell-based drug combination screening using an integrated droplet-based microfluidic system based on the sequential operation droplet array (SODA) technique. In the system, a tapered capillary connected with a syringe pump was used for multistep droplet manipulations. An oil-covered two-dimensional droplet array chip fixed in an x-y-z translation stage was used as the platform for cell culture and analysis. Complex multistep operations for drug combination screening involving long-term cell culture, medium changing, schedule-dependent drug dosage and stimulation, and cell viability testing were achieved in parallel in the semiopen droplet array, using multiple droplet manipulations including liquid metering, aspirating, depositing, mixing, and transferring. Long-term cell culture as long as 11 days was performed in oil-covered 500 nL droplets by changing the culture medium in each droplet every 24 h. The present system was applied in parallel schedule-dependent drug combination screening for A549 nonsmall lung cancer cells with the cell cycle-dependent drug flavopiridol and two anticancer drugs of paclitaxel and 5-fluorouracil. The highest inhibition efficiency was obtained with a schedule combination of 200 nM flavopiridol followed by 100 μM 5-fluorouracil. The drug consumption for each screening test was substantially decreased to 5 ng-5 μg, corresponding to 10-1000-fold reductions compared with traditional drug screening systems with 96-well or 384-well plates. The present work provides a novel and flexible droplet-based microfluidic approach for performing cell-based screening with complex and multistep operation procedures.
我们使用基于顺序操作液滴阵列（SODA）技术的集成液滴微流控系统进行基于细胞的药物组合筛选。在该系统中，使用带有注射器泵的锥形毛细管进行多步液滴操作。一个覆盖有油的二维液滴阵列芯片固定在 x-y-z 平移台上，用作细胞培养和分析的平台。复杂的多步药物组合筛选操作，包括长期细胞培养、介质更换、时间依赖性药物剂量和刺激以及细胞活力测试，在半开放式液滴阵列中通过多次液滴操作实现，包括液体计量、抽吸、沉积、混合和转移。通过每隔 24 小时更换每个液滴中的培养基，在覆盖有 500nL 油的液滴中进行长达 11 天的长期细胞培养。本系统应用于具有细胞周期依赖性药物 flavopiridol 和两种抗癌药物紫杉醇和 5-氟尿嘧啶的 A549 非小细胞肺癌细胞的并行时间依赖性药物组合筛选。在 200nM flavopiridol 后序贯使用 100μM 5-氟尿嘧啶的方案组合可获得最高的抑制效率。与 96 孔或 384 孔板的传统药物筛选系统相比，每个筛选测试的药物消耗大大减少至 5ng-5μg，减少了 10-1000 倍。本工作提供了一种新颖且灵活的基于液滴的微流控方法，用于进行具有复杂和多步操作程序的基于细胞的筛选。
