Analyzing Mechanisms of Metastatic Cancer Cell Adhesive Phenotype Leveraging Preparative Adhesion Chromatography Microfluidic

• Published in: Advanced biosystems 2019-03, Vol.3 (3), p.e1800328-n/a
• Main Authors: Birmingham, Katherine G., O'Melia, Meghan J., Ban, Dongjo, Mouw, Janna, Edwards, Erin E., Marcus, Adam I., McDonald, John, Thomas, Susan N.
• Format: Article
• Language: English
• Subjects: colon cancer; E‐selectin; metastasis; microfluidic; sialyl Lewis x
• ISSN: 2366-7478; 2366-7478
• DOI: 10.1002/adbi.201800328

An integrated, parallel‐plate microfluidic device is engineered to interrogate and fractionate cells based on their adhesivity to a substrate surface functionalized with adhesive ligand in a tightly controlled flow environment to elucidate associated cell‐intrinsic pathways. Wall shear stress levels and endothelial presentation of E‐selectin are modeled after the inflamed vasculature microenvironment in order to simulate in vitro conditions under which in vivo hematogenous metastasis occurs. Based on elution time from the flow channel, the collection of separate fractions of cells—noninteracting and interacting—at high yields and viabilities enables multiple postperfusion analyses, including flow cytometry, in vivo metastasis modeling, and transcriptomic analysis. This platform enables the interrogation of flow‐regulated cell molecular profiles, such as (co)expression levels of natively expressed selectin ligands sLex, CD44, and carcinoembryonic antigen, and cancer stem cell marker CD24. This additionally reveals E‐selectin adhesivity exhibited by metastatic human colon carcinoma cells to be a transient phenotype. Facile and rapid, this methodology for unbiased, label free sorting of large populations of cells based on their adhesion in flow represents a method of studying flow‐regulated adhesion in vitro for the identification of molecular drug targets for development as antimetastatic cancer therapeutics. In order to elucidate mechanisms of cancer cell metastasis, a parallel‐plate microfluidic flow chamber was engineered to interrogate and fractionate colon cancer cells based on their adhesivity to a substrate surface functionalized with adhesive ligands in physiological flow conditions. This platform enabled interrogation of cell molecular profiles that may underlie varying phenotypic characteristics of a heterogeneous metastatic cancer cell population.