Two-Step Acoustophoresis Separation of Live Tumor Cells from Whole Blood
Year: 2021
Type: article
Abstract: High Resolution Image Download MS PowerPoint Slide There is an unmet clinical need to extract living circulating tumor cells (CTCs) for functional studies and in vitro expansion to enable drug testing and predict responses to therapy in metastatic cancer. Here, we present a novel two-step acoustophoresis (A 2 ) method for isolation of unfixed, viable cancer cells from red blood cell (RBC) lysed whole blood. The A 2 method uses an initial acoustofluidic preseparation step to separate cells based on their acoustic mobility. This acoustofluidic step enriches viable cancer cells in a central outlet, but a significant number of white blood cells (WBCs) remain in the central outlet fraction due to overlapping acoustophysical properties of these viable cells. A subsequent purging step was employed to remove contaminating WBCs through negative selection acoustophoresis with anti-CD45-functionalized negative acoustic contrast particles. We processed 1 mL samples of 1:1 diluted RBC lysed whole blood mixed with 10 000 DU145 cells through the A 2 method. Additional experiments were performed using 1000 DU145 cells spiked into 1.5 × 10 6 WBCs in 1 mL of buffer to further elucidate the dynamic range of the method. Using samples with 10 000 DU145 cells, we obtained 459 ± 188-fold depletion of WBC and 42% recovery of viable cancer cells. Based on spiked samples with 1000 DU145 cells, our cancer cell recovery was 28% with 247 ± 156-fold WBC depletion corresponding to a depletion efficacy of ≥99.5%. The novel A 2 method provides extensive elimination of WBCs combined with the gentle recovery of viable cancer cells suitable for downstream functional analyses and in vitro culture.
Source: Analytical Chemistry
Institutions: Lund University, Memorial Sloan Kettering Cancer Center
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $5,000
Awards: G1324902806, G1456484, G2051006376, Ultrasonic standing wave fields and extra cellular vesicles enable novel routes in medical diagnostics and biopharmaceutical production, Acoustofluidic enrichment of extracellular vesicles for precision medicine in sepsis, G6062837801, G6265702644, G78939849, G8966693843, Cancer Center Support Grant, SPORE in Prostate Cancer
OpenAlex ID: W4200327042