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细胞培养监测在药物筛选和癌症研究中的应用:透明、微流控、多传感器微系统。

Cell culture monitoring for drug screening and cancer research: a transparent, microfluidic, multi-sensor microsystem.

机构信息

Laboratory for Sensors, Department of Microsystems Engineering (IMTEK), University of Freiburg, Georges-Köhler-Allee 103, 79110 Freiburg, Germany.

出版信息

Lab Chip. 2014 Jan 7;14(1):138-46. doi: 10.1039/c3lc50759a. Epub 2013 Nov 11.

Abstract

We present a novel, multiparametric microphysiometry system for the dynamic online monitoring of human cancer cell metabolism. The optically transparent, modular, hybrid microsystem is based on a glass chip and combines a cell cultivation chamber, microfluidics and metabolic monitoring with fully integrated chemo- and biosensors. pH and oxygen are measured in the cell culture area, and biosensors for lactate and glucose are connected downstream by microfluidics. The wafer-level fabrication features thin-film platinum and iridium oxide microelectrodes on a glass chip, microfluidics in an epoxy resist, a hybrid assembly and an on-chip reference electrode. The reliable analytical performance of the sensors in cell culture medium was demonstrated. The pH sensors exhibit a long-term stable, linear response. The oxygen sensors show a linear behaviour, which is also observed for low oxygen concentrations. Glucose and lactate measurements show a linear, long-term stable, selective and reversible behaviour in the desired range. T98G human brain cancer cells were cultivated and cell culture metabolism was measured on-chip. Stop/flow cycles were applied and extracellular acidification, respiration, glucose consumption and lactate production were quantified. Long-term metabolic rates were determined and all parameters could be measured in the outlet channel. A placement downstream of the cell cultivation area for biosensors was realised. A highly effective medium exchange and undiluted sampling from the cell culture chamber with low flow rates (2 μl min(-1)) and low volumes (15 μl per cycle) were achieved. The drug screening application was demonstrated by detecting alteration and recovery effects of cellular metabolism induced by the addition of substances to the medium.

摘要

我们提出了一种新颖的、多参数微生理测量系统,用于动态在线监测人类癌细胞代谢。该系统采用透明的、模块化的混合微系统,基于玻璃芯片,结合细胞培养室、微流控和代谢监测功能,以及完全集成的化学和生物传感器。在细胞培养区测量 pH 值和氧气,通过微流控将乳酸盐和葡萄糖生物传感器连接到下游。晶圆级制造技术在玻璃芯片上采用了薄膜铂和氧化铱微电极、环氧树脂微流控、混合组件和片上参考电极。传感器在细胞培养基中的可靠分析性能得到了证明。pH 传感器具有长期稳定、线性响应的特点。氧气传感器表现出线性行为,在低氧气浓度下也观察到这种行为。葡萄糖和乳酸测量在所需范围内表现出线性、长期稳定、选择性和可逆的行为。培养 T98G 人脑癌细胞并在芯片上测量细胞培养代谢。应用了停止/流动循环,并定量了细胞外酸化、呼吸、葡萄糖消耗和乳酸生成。确定了长期代谢率,并且所有参数都可以在出口通道中测量。在细胞培养区的下游实现了生物传感器的位置。通过以低流速(2 μl min(-1)) 和低体积(每个循环 15 μl)从细胞培养室进行高效的介质交换和未稀释采样,实现了这一目标。通过向培养基中添加物质来检测细胞代谢的改变和恢复效果,证明了该药物筛选应用。

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