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A hybrid microfluidic platform for cell-based assays via diffusive and convective trans-membrane perfusion

机译:一种混合微流体平台,用于通过扩散和对流跨膜灌注进行基于细胞的分析

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摘要

We present a novel 3D hybrid assembly of a polymer microfluidic chip with polycarbonate track-etched membrane (PCTEM) enabling membrane-supported cell culture. Two chip designs have been developed to establish either diffusive orudconvective reagent delivery using the integrated PCTEM. While it is well suited to a range of cell-based assays, we specifically employ this platform for the screening of a common antitumor chemotoxic agent (mitomycin C – MMC) on the HL60 myeloid leukemia cell line. The toxic activity of MMC is based on the generation of severe DNA damage in the cells. Using either mode of operation, the HL60 cells were cultured on-chip before, during, and after exposure to MMC at concentrations ranging from 0 to 50 lM. Cell viability was analysed off-chip by the trypan blue dye exclusion assay. The results of the on-chip viability assay were found to be consistent with those obtained off-chip and indicated ca. 40% cell survival at MMC concentration of 50 lM. The catalogue of capabilities of the here described cell assayudplatform comprises of (i) the culturing of cells either under shear-free conditions or under induced through-membrane flows, (ii) the tight time control of the reagentudexposure, (iii) the straightforward assembly of devices, (iv) the flexibility on the choice of the membrane, and, prospectively, (v) the amenability for large-scale parallelization.
机译:我们介绍了一种新型的3D混合组装的聚合物微流控芯片,具有聚碳酸酯轨迹蚀刻膜(PCTEM),可进行膜支持的细胞培养。已经开发出两种芯片设计,以使用集成式PCTEM建立扩散或非对流试剂输送。尽管它非常适用于一系列基于细胞的测定,但我们特别采用了该平台,用于筛选HL60髓样白血病细胞系中的常见抗肿瘤化学毒性剂(丝裂霉素C – MMC)。 MMC的毒性是基于细胞中严重DNA损伤的产生。使用任何一种操作模式,在暴露于MMC之前,之中和之后,均以0至50μM的浓度在芯片上培养HL60细胞。通过台盼蓝染料排除试验在芯片外分析细胞活力。发现片上生存力分析的结果与从片外获得的结果一致,并表明大约为1。在50μM的MMC浓度下,细胞存活率为40%。此处描述的细胞测定/平台的功能目录包括(i)在无剪切条件下或诱导的穿膜流下培养细胞,(ii)严格控制试剂/暴露时间,(iii )设备的直接组装;(iv)膜选择的灵活性;以及(v)大规模并行化的适应性。

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