[ACS Appl. Mater. Interfaces] Isolation of Phenotypically-Distinct Cancer Cells Using Nanoparticle-M
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 epithelial to mesenchymal transition (EMT). Here, we describe a two-dimensional separation approach that yields phenotypically-distinct subpopulations of cancer cells. Magnetic nanoparticles coated with antibodies against the epithelial cell adhesion molecule (EpCAM) are used to enrich cancer cells in a microfluidic platform. Cells are sorted into 4 zones based on the levels of EpCAM expression, which enables the detection of EMT- transformed cells. The aggressiveness of the isolated cells are then assessed using matrix- coated surfaces for collagen uptake analysis, and a separate dose- response NAD(PH) assay. SKBR3 breast cancer cells are sorted, and we find that low- EpCAM expressing cells have higher collagen uptake and higher folate-induced NAD(P)H responses compared to high-EpCAM expressing cells. Additionally, SKBR3 cells with chemically induced hypoxia (SKBR3-EMT cells) are used to mimic aggressive breast cancer. We find that SKBR3-EMT cells have higher collagen uptake and NAD(P)H metabolism relative to un-induced SKBR3 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.
Brenda Green, Leyla Kermanshah, Mahmoud Labib, Sharif Ahmed, Pamuditha Silva, Laili Mahmoudian, I-Hsin Chang, Reza M Mohamadi, Jonathan V. Rocheleau, and Shana O. Kelley ACS Appl. Mater. Interfaces, Just Accepted Manuscript DOI: 10.1021/acsami.7b05253 Publication Date (Web): May 26, 2017 Copyright © 2017 American Chemical Society