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Viscoelastic substrate decouples cellular traction force from other related phenotypes.

Biochemical and biophysical research communications (2021-01-29)
Nehal Dwivedi, Siddhartha Das, Jayesh Bellare, Abhijit Majumder
ABSTRACT

Survival and maintenance of normal physiological functions depends on continuous interaction of cells with its microenvironment. Cells sense the mechanical properties of underlying substrate by applying force and modulate their behaviour in response to the resistance offered by the substrate. Most of the studies addressing cell-substrate mechanical interactions have been carried out using elastic substrates. Since tissues within our body are viscoelastic in nature, here we explore the effect of substrate's viscoelasticity on various properties of mesenchymal stem cells. Here, we used two sets of polyacrylamide substrates having similar storage modulus (G' = 1.1-1.6 kPa) but different loss modulus (G" = 45 Pa and 300 Pa). We report that human mesenchymal stem cells spread more but apply less force on the viscoelastic substrate (substrate with higher loss modulus). We further investigated the effect of substrate viscoelasticity on the expression of other contractility-associated proteins such as focal adhesion (FA) proteins (Vinculin, Paxillin, Talin), cytoskeletal proteins (actin, mysion, intermediate filaments, and microtubules) and mechano-sensor protein Yes-Associated Protein (YAP). Our results show that substrate viscoelasticity decouples cellular traction from other known traction related phenotypes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Triton X-100, laboratory grade
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Monoclonal Anti-α-Tubulin antibody produced in mouse, clone DM1A, ascites fluid
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Monoclonal Anti-Vinculin antibody produced in mouse, clone hVIN-1, ascites fluid
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Monoclonal Anti-Talin antibody produced in mouse, clone 8d4, ascites fluid
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Anti-Myosin IIA, non muscle antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
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Monoclonal Anti-Vimentin antibody produced in mouse, clone VIM-13.2, ascites fluid