Abstract
PurposeWe aimed to generate a capture platform that integrates a deterministic lateral displacement (DLD) microfluidic structure with magnetic-activated cell sorting (MACS) technology for miniaturized, efficient, tumor antigen-independent circulating tumor cell (CTC) separation.MethodsThe microfluidic structure was based on the theory of DLD and was designed to remove most red blood cells and platelets. Whole Blood CD45 MicroBeads and a MACS separator were then used to remove bead-labeled white blood cells. We established HepG2 human liver cancer cells overexpressing green fluorescent protein by lentiviral transfection to simulate CTCs in blood, and these cells were then used to determine the CTC isolation efficiency of the device. The performance and clinical value of our platform were evaluated by comparison with the Abnova CytoQuest™ CR system in the separating of blood samples from 12 hepatocellular carcinoma patients undergoing liver transplantation in a clinical follow-up experiment. The isolated cells were stained and analyzed by confocal laser scanning microscopy.ResultsUsing our integrated platform at the optimal flow rates for the specimen (60 µl/min) and buffer (100 µl/min per chip), we achieved an CTC yield of 85.1% ± 3.2%. In our follow-up of metastatic patients, CTCs that underwent epithelial–mesenchymal transition were found. These CTCs were missed by the CytoQuest™ CR bulk sorting approach, whereas our platform displayed increased sensitivity to EpCAMlow CTCs.ConclusionsOur platform, which integrates microfluidic and MACS technology, is an attractive method for high-efficiency CTC isolation regardless of surface epitopes.
Highlights
Hepatocellular carcinoma (HCC) is one of the most common malignancies in the world (Omata et al, 2017)
Isolation efficiency and optimal flow rates for the platform integrating deterministic lateral displacement (DLD) with magnetic-activated cell sorting (MACS) separation HepG2-green fluorescent protein (GFP) cells were spiked into 10-fold diluted whole blood at a concentration of 1 Â 102 cells/ml
Liver transplantation is used for the complete removal of tumor lesions, and the tumor recurrence and metastasis observed in HCC patients is likely related to circulating tumor cell (CTC)
Summary
Hepatocellular carcinoma (HCC) is one of the most common malignancies in the world (Omata et al, 2017). A significant proportion of HCC patients who undergo LT still suffer from unfavorable outcomes due to the chance of tumor recurrence (Trojan, Zangos & Schnitzbauer, 2016). One reason for the poor prognosis of liver cancer is that cancer cells can enter and spread through the bloodstream. Before surgical resection, these circulating tumor cells (CTCs) may be present in the patient’s blood. CTCs are currently a major focus of liver cancer research because of the key roles they play in HCC recurrence and metastasis (Masuda et al, 2016; Kelley et al, 2015; Choi et al, 2015)
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