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Optimization design and drug analysis of cardiomyocyte based biosensor |
WANG Qin, FANG Jiaru, CAO Duanxi, ZHOU Jie,SU Kaiqi, LI Hongbo, WANG Ping |
Biosensor National Special Laboratory, Key Laboratory of Biomedical Engineering of Ministry of Education, Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, China |
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Abstract The coupling property between cardiomyocytes and microelectrode array (MEA) was studied from two aspects, including MEA surface hydrophilicity and cell culture density, in order to build a cardiomyocytebased potential biosensor of high stability and high consistency. MEA surface hydrophilicity was improved by high molecular protein (gelatin) modification, and the characteristics of extracellular field potential (EFP) signals of cardiomyocytebased potential biosensor under different cell densities were emphatically analyzed. Results show that the highclosely coupling between cardiomyocytes and MEA is formed after cardiomyocytes seeded on the MEA with gelatin modification at the proper cell density of 1.2×105 cells/cm2. Under the optimal conditions, the cardiomyocytebased potential biosensor presents highstable and highconsistent EFP signals, the spike amplitude (SA) reaches about 1.2 mV, and the firing rate (FR) reaches about 180 beats/min, which continues for 34 days. Two typical tool drugs, isoprotenol (ISO) and lidocaine (LID) were applied to test the analytical performance of the optimized cardiomyocytebased potential biosensor. Results show that SA and FR markedly increase after the treatment of 20 μM ISO for 5 min and significantly decrease after the treatment of 20 μM LID for 5 min. This biosensor performed rapid and sensitive response to these two drugs and will provide a promising platform for drug detection and analysis.
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Published: 31 December 2015
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心肌细胞传感器优化设计及其药物分析
为了构建高度稳定性和一致性的心肌细胞电位传感器,从微电极阵列表面亲水性和细胞培养密度两方面对心肌细胞和微电极阵列(MEA)的耦合性进行研究.通过对MEA表面进行高分子蛋白明胶的修饰来提高MEA表面的亲水性,并重点分析不同细胞密度下构建的心肌细胞电位传感器的胞外场电位信号(EFP)信号特征.研究结果表明:心肌细胞按优化密度12 万/cm2培养在经明胶修饰的MEA表面上,可促使心肌细胞和MEA形成高度紧密的耦合.在此条件下构建的心肌细胞传感器,能稳定输出一致性良好的EFP信号,电位幅值可达到约1.2 mV,发放频率可达到约180 次/min,信号稳定期可维持3~4 d.通过选择2种典型的工具药物异丙肾上腺素和利多卡因对优化后的心肌细胞电位传感器进行分析性能的测试,实验结果表明:20 μM的异丙肾上腺素和利多卡因分别大幅度增强和抑制了电位幅值和发放频率,结果与文献报导的结果相一致.该心肌细胞传感器对2种测试药物作出了快速而灵敏的响应,有望成为药物检测和分析的有效平台.
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