Two-stage microfluidic chip for selective isolation of circulating tumor cells (CTCs)
Two-stage microfluidic chip for selective isolation of circulating tumor cells (CTCs)
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DOI：
10.1016/j.bios.2014.07.019
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发表时间：
2015-05-15
影响因子：
12.6
通讯作者：
Jung, Hyo-Il
中科院分区：
文献类型：
--
作者：
Hyun, Kyung-A;Lee, Tae Yoon;Jung, Hyo-Il
在过去的几十年里，循环肿瘤细胞（CTC）一直被研究作为克服癌症的一种手段。然而，CTC 的稀有性和异质性一直是 CTC 研究的最大障碍。已经开发了许多用于 CTC 分离的技术，可分为正富集（即，利用细胞大小、表面蛋白表达等特异性分离靶细胞）和负富集（即，特异性洗脱非靶细胞）。正富集方法纯度较高，但可能因其选择标准而产生偏差，而负富集方法纯度相对较低，但可以分离异质 CTC。为了弥补正负富集的已知缺点，在本研究中我们引入了两级微流控芯片。第一阶段涉及微流控磁激活细胞分选 (mu-MACS) 芯片来洗脱白细胞 (WBC)。第二阶段涉及几何激活表面相互作用 (GASI) 芯片，用于选择性分离 CTC。我们观察到使用 mu-MACS 芯片以 400 μL/min 的流速掺入 5 mL 血液样本中的癌细胞富集度高达 763 倍。使用 GASI 芯片以 100 μL/min 的流速成功分离癌细胞，基于 EpCAM 或 HER2 表面蛋白表达，分离效率为 10.19% 至 22.91%。我们的两级微流控芯片不仅可以从血细胞中分离出CFC，还可以根据其特征对异质CFC进行分类。因此，我们的芯片可以有助于 CTC 异质性的研究，进而有助于个性化癌症治疗。 (C) 2014 Elsevier B.V. 保留所有权利。
Over the past few decades, circulating tumor cells (CTCs) have been studied as a means of overcoming cancer. However, the rarity and heterogeneity of CTCs have been the most significant hurdles in CTC research. Many techniques for,CTC isolation have been developed and can be classified into positive enrichment (i.e., specifically isolating target cells using cell size, surface protein expression, and so on) and negative enrichment (i.e., specifically eluting non-target cells). Positive enrichment methods lead to high purity, but could be biased by their selection criteria, while the negative enrichment methods have relatively low purity, but can isolate heterogeneous CTCs. To compensate for the known disadvantages of the positive and negative enrichments, in this study we introduced a two-stage microfluidic chip. The first stage involves a microfluidic magnetic activated cell sorting (mu-MACS) chip to elute white blood cells (WBCs). The second stage involves a geometrically activated surface interaction (GASI) chip for the selective isolation of CTCs. We observed up to 763-fold enrichment in cancer cells spiked into 5 mL of blood sample using the mu-MACS chip at 400 mu L/min flow rate. Cancer cells were successfully separated with separation efficiencies ranging from 10.19% to 22.91% based on their EpCAM or HER2 surface protein expression using the GASI chip at a 100 mu L/min flow rate. Our two-stage microfluidic chips not only isolated CFCs from blood cells, but also classified heterogeneous CFCs based on their characteristics. Therefore, our chips can contribute to research on CTC heterogeneity of CTCs, and, by extension, personalized cancer treatment. (C) 2014 Elsevier B.V. All rights reserved.