Isolation of Phenotypically Distinct Cancer Cells Using Nanoparticle-Mediated Sorting

Abstract

Isolating subpopulations of heterogeneous cancer cells is an important capability for the meaningful characterization of circulating tumor cells at different stages of tumor progression and during the epithelial-to-mesenchymal transition. Here, we present a microfluidic device that can separate phenotypically distinct subpopulations of cancer cells. Magnetic nanoparticles coated with antibodies against the epithelial cell adhesion molecule (EpCAM) are used to separate breast cancer cells in the microfluidic platform. Cells are sorted into different zones on the basis of the levels of EpCAM expression, which enables the detection of cells that are losing epithelial character and becoming more mesenchymal. The phenotypic properties of the isolated cells with low and high EpCAM are then assessed using matrix-coated surfaces for collagen uptake analysis, and an NAD(P)H assay that assesses metabolic activity. We show that low-EpCAM expressing cells have higher collagen uptake and higher folate-induced NAD(P)H responses compared to those of high-EpCAM expressing cells. In addition, we tested SKBR3 cancer cells undergoing chemically induced hypoxia. The induced cells have reduced expression of EpCAM, and we find that these cells have higher collagen uptake and NAD(P)H metabolism relative to noninduced cells. This work demonstrates that nanoparticle-mediated binning facilitates the isolation of functionally distinct cell subpopulations and allows surface marker expression to be associated with invasiveness, including collagen uptake and metabolic activity.

Finding the perfect match between nanoparticles and microfluidics to respond to cancer challenges

“…Noteworthy, more recently, the same group published other studies where they showed that cells with low EpCAM expression were related with disease progression since they presented higher collagen uptake and higher metabolic activity, for so, a more invasive behaviour, than cells with higher EpCAM expression ( Figure 3, B). 45 As well, they showed that patients were less responsive to therapy presented low EpCAM expression and high levels of cancer biomarkers, such as androgen receptor variant 7 (ARV7) and the mesenchymal marker, Ncadherin. 46 Despite the advance achieved in CTCs capture by using magnetic approaches, some issues still need to be addressed, such as nanoparticle aggregation, and consequently, cell aggregation.…”

Detection of Rare Objects by Flow Cytometry: Imaging, Cell Sorting, and Deep Learning Approaches

“…Huang et al designed a microfluidic microwell device that allows for immunomagnetic single-cell trapping for further cellular analysis at a single-cell level [238]. Green et al described a microfluidic magnetic sorting device that allows for the effective capturing of CTCs and dividing them into subpopulations according to levels of protein surface expression [248]. The analysis of subpopulations of CTCs that exhibit different biochemical and functional phenotypes may be useful to figure out if the CTCs acquire the metastatic potential by undergoing the epithelial-to-mesenchymal transition (EMT).…”

Magnetic Ranking Cytometry: Profiling Rare Cells at the Single-Cell Level

“…Second-gen MagRC Number of zones/outlets 1 zone 1 4 zones 2, 3 6 zones 4 20 zones 5 8 zones 6, 7 10 zones 8 100 zones 9 3 outlets 10…”