Selective isolation of magnetic nanoparticle-mediated heterogeneity subpopulation of circulating tumor cells using magnetic gradient based microfluidic system

Abstract

Relocation mechanisms of the circulating tumor cells (CTCs) from the primary site to the secondary site through the blood vessel network cause tumor metastasis. Despite of the importance to diagnose the cancer metastasis by CTCs, still it is formidable challenge to use in the clinical purpose because of the rarity and the heterogeneity of CTCs in the cancer patient's peripheral blood sample. In this study we have developed magnetic force gradient based microfluidic chip (Mag-Gradient Chip) for isolating the total number of CTCs in the sample and characterizing the state of CTCs simultaneously with respect to the epithelial cell adhesion molecule (EpCAM) expression level. We have synthesized magnetic nanoparticles (MNPs) using hydrothermal method and functionalized anti-EpCAM on their surface for the specific binding with CTCs. The Mag-Gradient Chip designed to isolate and classify the CTCs by isolating at the different location in the chip using magnetic force differences depending on the EpCAM expression level. We observed 95.7% of EpCAM positive and 79.3% of EpCAM negative CTCs isolated in the Mag-Gradient Chip. At the same time, the 71.3% of isolated EpCAM positive CTCs were isolated at the first half area whereas the 76.9% of EpCAM negative CTCs were collected at the latter half area. The Mag-Gradient Chip can isolate the 3ml of heterogeneous CTCs sample in 1h with high isolating yield. The EpCAM expression level dose not means essential condition of the metastatic CTCs, but the Mag-Gradient Chip can shorten the date to diagnose the cancer metastasis in clinic.

Keywords: Circulating tumor cells (CTCs); Epithelial cell adhesion molecule (EpCAM); Heterogeneity; Magnetic activated cell sorting (MACS); Magnetic gradient chip (Mag-Gradient Chip).