Abstract:
This study focuses on the use of pulsed electric fields (PEF) in microfluidics for controlled cell studies. The commonly used material for soft lithography, polydimethylsiloxane (PDMS), does not fully ensure the necessary chemical and mechanical resistance in these systems. Integration of specific analytical measurement setups into microphysiological systems (MPS) are also challenging. We present an off-stoichiometry thiol-ene (OSTE)-based microchip, containing integrated electrodes for PEF and transepithelial electrical resistance (TEER) measurement and the equipment to monitor pH and oxygen concentration in situ. The effectiveness of the MPS was empirically demonstrated through PEF treatment of the C6 cells. The effects of PEF treatment on cell viability and permeability to the fluorescent dye DapI were tested in two modes: stop flow and continuous flow. The maximum permeability was achieved at 1.8 kV/cm with 16 pulses in stop flow mode and 64 pulses per cell in continuous flow mode, without compromising cell viability. Two integrated sensors detected changes in oxygen concentration before and after the PEF treatment, and the pH shifted towards alkalinity following PEF treatment. Therefore, our proof-of-concept technology serves as an MPS for PEF treatment of mammalian cells, enabling in situ physiological monitoring.
摘要:
这项研究的重点是在微流控细胞研究中使用脉冲电场(PEF)。常用的软光刻材料,聚二甲基硅氧烷(PDMS),不能完全确保这些系统中必要的耐化学性和耐机械性。将特定的分析测量设置集成到微生理系统(MPS)中也具有挑战性。我们提出了一种基于非化学计量的硫醇-烯(OSTE)的微芯片,包含用于PEF和跨上皮电阻(TEER)测量的集成电极以及用于原位监测pH和氧气浓度的设备。MPS的有效性通过C6细胞的PEF处理经验证明。在两种模式下测试PEF处理对细胞活力和对荧光染料DapI的渗透性的影响:停止流动和连续流动。最大磁导率在1.8kV/cm下实现,停流模式下16个脉冲,连续流模式下每个电池64个脉冲。不损害细胞活力。两个集成传感器检测到PEF治疗前后氧气浓度的变化,PEF处理后,pH向碱度移动。因此,我们的概念验证技术作为哺乳动物细胞PEF治疗的MPS,实现原位生理监测。