School of Mechanical Engineering, and, Jiangsu Key Laboratory for Design and Manufacture of Micro-Nano Biomedical Instruments, Southeast University, Nanjing, 211189, China.
Department of Oncology, Zhongda Hospital, Southeast University, Nanjing, 210009, China.
Lab Chip. 2022 May 31;22(11):2097-2106. doi: 10.1039/d2lc00082b.
Accurate and rapid diagnosis of malignant pleural and peritoneal effusions is critical due to potential association with advanced disease stages or progression. Traditional cytodiagnosis suffers from low efficiency and has difficulties in finding malignant tumor cells (MTCs) from a mass of exfoliated cells. Hence, a polymer microfluidic chip with a slanted spiral channel was employed for high-throughput and label-free enrichment of MTCs and MTC clusters from clinical malignant pleural and peritoneal effusions. The slanted spiral channel with trapezoidal cross-sections was fabricated by assembling two patterned polymer films of different thicknesses within one flow channel layer. After systematically exploring the effects of the particle size, effusion concentration, and flow rate on separation performance of the device, we realized the enrichment of MTCs from abundant blood cells in 2-fold diluted effusions. The results indicated that approximately 85% of the spiked tumor cells (A549 and MCF-7 cell lines) were recovered with high purities of over 37% at a high throughput of 2000 μL min. In clinical applications, we successfully enriched 24-2691 MTCs per mL from the diluted malignant pleural and peritoneal effusions collected from four types of cancer patients ( = 22). More importantly, the MTC clusters were further purified from single MTCs using a higher flow rate of 3000 μL min. Finally, we performed the rapid drug sensitivity test by coupling the microfluidic enrichment with CCK-8 assay. Our approach may serve as valuable assistance to accelerate cancer diagnosis and guide the selection of treatment medications.
准确快速地诊断恶性胸腔和腹腔积液至关重要，因为这可能与晚期疾病阶段或进展有关。传统的细胞学诊断效率低下，难以从大量脱落细胞中找到恶性肿瘤细胞（MTCs）。因此，采用具有倾斜螺旋通道的聚合物微流控芯片，从临床恶性胸腔和腹腔积液中高通量、无标记地富集 MTC 和 MTC 簇。具有梯形横截面的倾斜螺旋通道是通过在一个流道层内组装两个不同厚度的图案化聚合物薄膜来制造的。在系统地研究了粒径、积液浓度和流速对器件分离性能的影响后，我们实现了从 2 倍稀释的积液中丰富的血细胞中富集 MTC。结果表明，大约 85%的掺入肿瘤细胞（A549 和 MCF-7 细胞系）以超过 37%的高纯度在 2000 μL min 的高通量下回收。在临床应用中，我们成功地从四种癌症患者的稀释恶性胸腔和腹腔积液中富集了每毫升 24-2691 个 MTC（n = 22）。更重要的是，通过将微流控富集与 CCK-8 测定相结合，我们从单个 MTC 中进一步纯化了 MTC 簇。最后，我们通过将微流控富集与 CCK-8 测定相结合，进行了快速药物敏感性测试。我们的方法可以为加速癌症诊断和指导治疗药物选择提供有价值的帮助。
