High-throughput and label-free enrichment of malignant tumor cells and clusters from pleural and peritoneal effusions using inertial microfluidics

Abstract

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⁻¹. 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 (n = 22). More importantly, the MTC clusters were further purified from single MTCs using a higher flow rate of 3000 μL min⁻¹. 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.