AI Article Synopsis
- Cellular contractility plays a crucial role in various biological functions, from cell movement and division to maintaining tissue stability, and is crucial for the functioning of multi-cellular organisms.
- Disruptions in contractility are linked to various diseases, making it a valuable target for new diagnostic techniques and therapeutic approaches, which require accurate measurement of contractile forces.
- The study introduces a novel method using silicone elastomer-based traction force microscopy in a multi-well format to analyze the effects of TGF-β on contractility during the Epithelial to Mesenchymal Transition (EMT) in NMuMG cells, highlighting its relevance in understanding disease mechanisms.
Article Abstract
Cellular contractility is essential in diverse aspects of biology, driving processes that range from motility and division, to tissue contraction and mechanical stability, and represents a core element of multi-cellular animal life. In adherent cells, acto-myosin contraction is seen in traction forces that cells exert on their substrate. Dysregulation of cellular contractility appears in a myriad of pathologies, making contractility a promising target in diverse diagnostic approaches using biophysics as a metric. Moreover, novel therapeutic strategies can be based on correcting the apparent malfunction of cell contractility. These applications, however, require direct quantification of these forces. We have developed silicone elastomer-based traction force microscopy (TFM) in a parallelized multi-well format. Our use of a silicone rubber, specifically polydimethylsiloxane (PDMS), rather than the commonly employed hydrogel polyacrylamide (PAA) enables us to make robust and inert substrates with indefinite shelf-lives requiring no specialized storage conditions. Unlike pillar-PDMS based approaches that have a modulus in the GPa range, the PDMS used here is very compliant, ranging from approximately 0.4 kPa to 100 kPa. We create a high-throughput platform for TFM by partitioning these large monolithic substrates spatially into biochemically independent wells, creating a multi-well platform for traction force screening that is compatible with existing multi-well systems. In this manuscript, we use this multi-well traction force system to examine the Epithelial to Mesenchymal Transition (EMT); we induce EMT in NMuMG cells by exposing them to TGF-β, and to quantify the biophysical changes during EMT. We measure the contractility as a function of concentration and duration of TGF-β exposure. Our findings here demonstrate the utility of parallelized TFM in the context of disease biophysics.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8033630 | PMC |
| http://dx.doi.org/10.3791/59364 | DOI Listing |
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