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利用介电泳对微流控剪切依赖性上皮细胞黏附分子免疫捕获和从血细胞中富集胰腺癌细胞的特性研究。

Characterization of microfluidic shear-dependent epithelial cell adhesion molecule immunocapture and enrichment of pancreatic cancer cells from blood cells with dielectrophoresis.

机构信息

Department of Biomedical Engineering, Cornell University , Ithaca, New York 14853, USA.

Sibley School of Mechanical and Aerospace Engineering, Cornell University , Ithaca, New York 14853, USA.

出版信息

Biomicrofluidics. 2014 Jul 21;8(4):044107. doi: 10.1063/1.4890466. eCollection 2014 Jul.

Abstract

Current microfluidic techniques for isolating circulating tumor cells (CTCs) from cancer patient blood are limited by low capture purity, and dielectrophoresis (DEP) has the potential to complement existing immunocapture techniques to improve capture performance. We present a hybrid DEP and immunocapture Hele-Shaw flow cell to characterize DEP's effects on immunocapture of pancreatic cancer cells (Capan-1, PANC-1, and BxPC-3) and peripheral blood mononuclear cells (PBMCs) with an anti-EpCAM (epithelial cell adhesion molecule) antibody. By carefully specifying the applied electric field frequency, we demonstrate that pancreatic cancer cells are attracted to immunocapture surfaces by positive DEP whereas PBMCs are repelled by negative DEP. Using an exponential capture model to interpret our capture data, we show that immunocapture performance is dependent on the applied DEP force sign and magnitude, cell surface EpCAM expression level, and shear stress experienced by cells flowing in the capture device. Our work suggests that DEP can not only repel contaminating blood cells but also enhance capture of cancer cell populations that are less likely to be captured by traditional immunocapture methods. This combination of DEP and immunocapture techniques to potentially increase CTC capture purity can facilitate subsequent biological analyses of captured CTCs and research on cancer metastasis and drug therapies.

摘要

当前从癌症患者血液中分离循环肿瘤细胞 (CTC) 的微流控技术受到低捕获纯度的限制,而电泳(DEP)有可能补充现有的免疫捕获技术,以提高捕获性能。我们提出了一种混合 DEP 和免疫捕获的 Hele-Shaw 流控芯片,用于表征 DEP 对用抗 EpCAM(上皮细胞黏附分子)抗体进行免疫捕获的胰腺癌细胞(Capan-1、PANC-1 和 BxPC-3)和外周血单核细胞(PBMCs)的影响。通过仔细指定施加的电场频率,我们证明胰腺癌细胞通过正 DEP 被吸引到免疫捕获表面,而 PBMCs 则被负 DEP 排斥。我们使用指数捕获模型来解释我们的捕获数据,结果表明免疫捕获性能取决于施加的 DEP 力的符号和大小、细胞表面 EpCAM 的表达水平以及在捕获装置中流动的细胞所经历的剪切应力。我们的工作表明,DEP 不仅可以排斥污染的血细胞,还可以增强对传统免疫捕获方法不太可能捕获的癌细胞群体的捕获。DEP 和免疫捕获技术的这种组合有可能提高 CTC 的捕获纯度,从而促进对捕获的 CTC 进行后续的生物学分析以及对癌症转移和药物治疗的研究。

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