A high-throughput, open-space and reusable microfluidic chip for combinational drug screening on tumor spheroids
A high-throughput, open-space and reusable microfluidic chip for combinational drug screening on tumor spheroids
复制标题
一种高通量、开放空间、可重复使用的微流控芯片，用于肿瘤球体的组合药物筛选
DOI：
10.1039/d1lc00525a
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发表时间：
2021-08-24
期刊：
影响因子：
6.1
通讯作者：
Huang, Wei
中科院分区：
文献类型：
--
作者：
Li, Lijun;Chen, Yan;Huang, Wei
利用肿瘤球体筛选药物组合对疾病治疗和个体化用药的发展具有重要作用。然而，目前的研究大多集中在药物梯度或两种药物的组合，很难找到涉及更多药物的复杂治疗组合。实验设计（DOE）微流体的使用是系统研究这一领域的潜在策略。在这里，我们开发了一个高通量，开放空间的多层PMMA微流控芯片的组合药物筛选肿瘤球体。这种微芯片制造简单，与标准球体培养物兼容，对最终用户友好。该装置由入口层和多个分散层组成。在入口层中，不同的样本可以同时加载到芯片中。样品溶液流入分散层，根据特定的DOE原理产生各种组合。我们证明了芯片的性能与设计定量一致，使用水和强力霉素组合作为模型。作为概念验证研究，我们构建了HeLa报告细胞系来量化肿瘤球体的自噬，并使用芯片来识别与球体生长相关的关键因素。具体而言，我们使用L-谷氨酰胺，D-葡萄糖，FBS和顺铂作为因子，并研究了自噬，生长曲线和球体大小对四种因子的不同组合的响应。我们发现d-葡萄糖可以抑制顺铂对肿瘤球体的作用，与2D单培养细胞相比，顺铂在3D肿瘤球体中引起严重的自噬。我们的方法有可能允许更多的药物组合进行检查，它可以扩展到DOE方法与七个或更多的输入。
Screening drug combinations using tumor spheroids can play a vital role in the development of disease treatment and personalized medicine. However, current studies focus on drug gradients or combinations of two drugs in most cases, and it is difficult to find complex therapeutic combinations involving more drugs. The use of design-of-experiment (DOE) microfluidics is a potential strategy to study this area systematically. Here we develop a high-throughput, open-space multilayered PMMA microfluidic chip for combinational drug screening on tumor spheroids. This microchip is straightforward to fabricate, compatible with standard spheroid cultures, and friendly for end-users. The device consists of an inlet layer and multiple dispersing layers. In the inlet layer, different samples can be loaded into the chip simultaneously. The sample solutions flow into the dispersing layers to generate various combinations based on the specific DOE principle. We demonstrated that the chip performance is in quantitative agreement with the design, using water and doxycycline combinations as models. As a proof-of-concept study, we constructed a HeLa reporter cell line to quantify the autophagy of tumor spheroids and used the chip to identify critical factors relating to the growth of the spheroids. Specifically, we used l-glutamine, d-glucose, FBS, and cisplatin as the factors and studied the autophagy, growth curves, and spheroid sizes in response to different combinations of the four factors. We found that d-glucose can inhibit the effects of cisplatin on tumor spheroids, and cisplatin caused severe autophagy in 3D tumor spheroids compared to 2D monoculture cells. Our method has the potential to allow more drug combinations to be examined, and it can be extended to DOE approaches with seven or more inputs.