Using digital holographic microscopy (DHM) to monitor effects of extracellular matrix (ECM) glycation on cancer cell morphology and migration

Abstract

The extracellular matrix (ECM) is a complex network of ubiquitously present acellular material that plays a critical role in cell proliferation, migration, invasion, and tissue morphogenesis. Non-enzymatic glycation of ECM modifies the structure and function of ECM proteins and can support a pro-inflammatory milieu in the tumor microenvironment. However, the impact of glycated ECM on cancer cell growth remains underexplored despite its importance in facilitating disease progression. Here, we investigate the effect of ECM glycation on cancer cell morphology and migration behavior. We used methylglyoxal (MG) as a glycation agent and collagen as our ECM model protein. For in vitro growth analysis, breast cancer cells were seeded on growth surfaces coated with both non-glycated and glycated collagen. Cell behavior was monitored for 24 hours using a real-time holographic imaging system. Holographic image analysis revealed significant differences between non-glycated and glycated growth substrates in cell spreading area, eccentricity, perimeter length, optical thickness, and optical volume, as well as cell migration and motility, which directly influence cell adhesion and proliferation. These changes were found to be cell line biased. Overall, our findings suggest that ECM glycation has a significant effect on cell morphology, migration and cell growth. Holographic live cell imaging was determined to be an excellent method to monitor cells without the need for any labeling and with minimal perturbations.