Cascaded filter deterministic lateral displacement microchips for isolation and molecular analysis of circulating tumor cells and fusion cells
Cascaded filter deterministic lateral displacement microchips for isolation and molecular analysis of circulating tumor cells and fusion cells
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用于循环肿瘤细胞和融合细胞的分离和分子分析的级联过滤器确定性横向位移微芯片
DOI：
10.1039/d1lc00360g
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发表时间：
2021-06-28
期刊：
影响因子：
6.1
通讯作者：
Chen, Yan
中科院分区：
文献类型：
--
作者：
Liu, Zongbin;Huang, Yuqing;Chen, Yan
从血液样本中精确分离和分析循环肿瘤细胞（CTC）为癌症研究和个性化治疗提供了相当大的潜力。目前，可用的CTC分离方法在寻求实现具有高分离效率和高纯度的细胞分离的简单策略方面仍然具有挑战性，这限制了捕获的CTC用于下游分析的使用。在这里，我们提出了一个过滤器确定性的横向位移的概念，以实现一步和无标记的CTC隔离与高通量。与传统的确定性横向位移（DLD）装置不同，所提出的方法通过将过滤概念结合到DLD结构中来使用流体动力学细胞分选设计，并通过级联微流体设计实现高通量和无堵塞的分离。级联过滤器-DLD（CFD）设计证明了基于尺寸的细胞分离的增强性能，并且实现了高分离效率（>96%）、高细胞纯度（WBC去除率99.995%）、高细胞活力（>98%）和高处理速率（lmL min（-1））。使用CFD-Chip分析来自肺癌患者的样品，有效地收集CTC和肿瘤细胞-白细胞融合细胞，并且CTC水平的变化用于治疗反应监测。CFD-Chip平台分离出具有良好活力的CTC，从而能够通过单细胞RNA测序进行直接下游分析。富集的CTC的转录组分析鉴定了CTC的新亚型，例如肿瘤细胞-白细胞融合细胞，从而为癌症诊断和治疗提供了见解。
Precise isolation and analysis of circulating tumor cells (CTCs) from blood samples offer considerable potential for cancer research and personalized treatment. Currently, available CTC isolation approaches remain challenging in the quest for simple strategies to achieve cell isolation with both high separation efficiency and high purity, which limits the use of captured CTCs for downstream analyses. Here, we present a filter deterministic lateral displacement concept to achieve one-step and label-free CTC isolation with high throughput. Unlike conventional deterministic lateral displacement (DLD) devices, the proposed method uses a hydrodynamic cell sorting design by incorporating a filtration concept into a DLD structure, and enables high-throughput and clog-free isolation by a cascaded microfluidic design. The cascaded filter-DLD (CFD) design demonstrated enhanced performance for size-based cell separation, and achieved high separation efficiency (>96%), high cell purity (WBC removal rate 99.995%), high cell viability (>98%) and high processing rate (1 mL min(-1)). Samples from lung cancer patients were analyzed using the CFD-Chip, CTCs and tumor cell-leukocyte fusion cells were efficiently collected, and changes in CTC levels were used for treatment response monitoring. The CFD-Chip platform isolated CTCs with good viability, enabling direct downstream analysis with single-cell RNA sequencing. Transcriptome analysis of enriched CTCs identified new subtypes of CTCs such as tumor cell-leukocyte fusion cells, providing insights into cancer diagnostics and therapeutics.