Elastic moduli of living epithelial pancreatic cancer cells and their skeletonized keratin intermediate filament network.

ABSTRACT

In simple epithelia, such as living epithelial pancreatic cancer cells (Panc-1), unusual amounts of keratin filaments can be found, which makes these cells an ideal model system to study the role of keratin for cell mechanical properties. In this work, the elastic moduli of Panc-1 cells and their extracted in-situ subcellular keratin intermediate filament network are determined and compared with each other. For this, the living adherent cells and their extracted keratin network were probed with local quasistatic indentation testing during large deformations using the Atomic Force Microscope (AFM). We determined the elastic modulus of the skeletonized but structurally intact keratin network to be in the order of 10 Pa, while the living cell elastic modulus ranged from 100 to 500 Pa. By removing microfilaments, microtubules, membranes and soluble cytoplasmic components during keratin network extraction, we excluded effects caused by crosslinking with other filamentous fibers and from the viscosity of the cytoplasm. Thus, the determined elastic modulus equals the actual elastic modulus inherent to such a keratin filamentous network. In our assessment of the effective mechanical contribution of the architecturally intact, skeletonized keratin network to living cell mechanics, we come to the conclusion that it plays only a very limited role. Evidently, the quantitative dominance of keratin in these cells does not reflect a strong influence on determining the cell's elastic modulus. Instead, keratin like other filamentous structures in the cell's scaffolding, e.g., F-actin and microtubuli, is one part of a greater whole.