Centrifugal microfluidic platform for single-cell level cardiomyocyte-based drug profiling and screening
Centrifugal microfluidic platform for single-cell level cardiomyocyte-based drug profiling and screening
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DOI：
10.1039/c5lc00652j
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发表时间：
2015-01-01
期刊：
影响因子：
6.1
通讯作者：
Tamiya, E.
中科院分区：
文献类型：
--
作者：
Espulgar, W.;Aoki, W.;Tamiya, E.
药物筛选和分析是药物发现、开发和上市的重要阶段。然而，由于需要额外的技术技能和昂贵的维护费用，一些图谱测试并不是常规进行的，这会导致意外副作用或药物不良反应(ADR)的情况。本研究介绍了一种用于单细胞水平药物筛选和图谱的微流控芯片的设计和运行，作为实现这一目的的替代平台。用离心法将分离的单个和多组原代培养的新生大鼠心肌细胞捕获在同一芯片中。在芯片的脱离旋转操作中，可以在显微镜下观察细胞，并可以记录节拍运动的电影。通过对记录的视频进行图像相关分析，生成细胞的搏动曲线，以研究细胞的收缩特性(搏动频率、搏动强度和搏动间歇时间)。通过使用这种非侵入性工具，可以在捕获后立即进行长期连续监测，并成功地在芯片中观察到细胞生长和动态。介质和液体的替换不需要进一步的离心法，而是仅使用毛细管流。证明了卡巴胆碱(100 MU M)和异丙肾上腺素(4 mU g m L(-1))对单个细胞和多组细胞的影响，并揭示了该芯片的免疫染色(β-肌动蛋白)适用性。此外，这些发现可能有助于非侵入性心肌细胞图谱的进展，以及未来高通量芯片上实验室设备的芯片设计和操作。
Drug screening and profiling is an important phase in drug discovery, development, and marketing. However, some profiling tests are not routinely done because of the needed additional technical skills and costly maintenance, which leads to cases of unexpected side effects or adverse drug reactions (ADRs). This study presents the design and operation of a microfluidic chip for single-cell level drug screening and profiling as an alternative platform for this purpose. Centrifugation was utilized to trap isolated single and groups of primary cultured neonatal rat cardiomyocytes in the same chip. In the off-spin operation of the chip, the cells can be observed under a microscope and movies of the beat motion can be recorded. The beat profiles of the cells were generated by image correlation analysis of the recorded video to study the contractile characteristics (beating rate, beating strength, and inter-beat duration). By utilizing this non-invasive tool, long term continuous monitoring, right after trapping, was made possible and cell growth and dynamics were successfully observed in the chip. Media and liquid replacement does not require further centrifugation but instead utilizes capillary flow only. The effect of carbachol (100 mu M) and isoproterenol (4 mu g mL(-1)) on single cells and groups of cells was demonstrated and the feature for immunostaining (beta-actin) applicability of the chip was revealed. Furthermore, these findings can be helpful for the headway of non-invasive profiling of cardiomyocytes and for future chip design and operation of high-throughput lab-on-a-chip devices.