Department of Chemistry, The University of Georgia, Athens, Georgia, USA.
FCS Technology, LLC, Athens, GA, 30606, USA.
Lab Chip. 2021 Sep 14;21(18):3583-3597. doi: 10.1039/d1lc00454a.
Profiling circulating tumour cells (CTCs) in cancer patients' blood samples is critical to understand the complex and dynamic nature of metastasis. This task is challenged by the fact that CTCs are not only extremely rare in circulation but also highly heterogeneous in their molecular programs and cellular functions. Here we report a combinational approach for the simultaneous biochemical and functional phenotyping of patient-derived CTCs, using an integrated inertial ferrohydrodynamic cell separation (iFCS) method and a single-cell microfluidic migration assay. This combinatorial approach offers unique capability to profile CTCs on the basis of their surface expression and migratory characteristics. We achieve this using the iFCS method that successfully processes whole blood samples in a tumor cell marker and size agnostic manner. The iFCS method enables an ultrahigh blood sample processing throughput of up to 2 × 10 cells s with a blood sample flow rate of 60 mL h. Its short processing time (10 minutes for a 10 mL sample), together with a close-to-complete CTC recovery (99.70% recovery rate) and a low WBC contamination (4.07-log depletion rate by removing 99.992% of leukocytes), results in adequate and functional CTCs for subsequent studies in the single-cell migration device. For the first time, we employ this new approach to query CTCs with single-cell resolution in accordance with their expression of phenotypic surface markers and migration properties, revealing the dynamic phenotypes and the existence of a high-motility subpopulation of CTCs in blood samples from metastatic lung cancer patients. This method could be adopted to study the biological and clinical value of invasive CTC phenotypes.
对癌症患者血液样本中的循环肿瘤细胞（CTC）进行分析，对于了解转移的复杂和动态性质至关重要。然而，CTC 在血液循环中不仅极其罕见，而且在分子程序和细胞功能上也具有高度异质性，这给这项任务带来了挑战。在这里，我们报告了一种组合方法，用于通过集成惯性铁磁流体动力学细胞分离（iFCS）方法和单细胞微流控迁移分析来对患者来源的 CTC 进行同时的生化和功能表型分析。这种组合方法提供了基于其表面表达和迁移特性对 CTC 进行分析的独特能力。我们使用 iFCS 方法来实现这一点，该方法以肿瘤细胞标志物和大小不可知的方式成功处理全血样本。iFCS 方法能够以高达 2×10 个细胞/s 的超高血液样本处理通量和 60 mL/h 的血液样本流速进行处理。其处理时间短（10 分钟处理 10 毫升样本），接近完全的 CTC 回收率（99.70%的回收率）和低白细胞污染（通过去除 99.992%的白细胞，白细胞减少率为 4.07 个对数），为后续在单细胞迁移设备中的研究提供了足够且功能正常的 CTC。我们首次采用这种新方法，根据其表面表型标志物和迁移特性，以单细胞分辨率对 CTC 进行查询，揭示了转移性肺癌患者血液样本中 CTC 的动态表型和高迁移亚群的存在。该方法可用于研究侵袭性 CTC 表型的生物学和临床价值。
