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Observing and Quantifying Fibroblast-mediated Fibrin Gel Compaction
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Force fluctuations in three-dimensional suspended fibroblasts.

Florian Schlosser1, Florian Rehfeldt2, Christoph F Schmidt3

  • 1Third Institute of Physics-Biophysics, Georg August University, 37077 Göttingen, Germany.

Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
|December 24, 2014
PubMed
Summary

Cells generate forces influencing their environment, measured using optical traps. Myosin II activity, not microtubules, dictates cell stiffness and force generation, revealing insights into cellular mechanics.

Keywords:
active mattercell cortexcell mechanicsnon-muscle myosin IIoptical trap

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Area of Science:

  • Cellular mechanics
  • Biophysics
  • Cytoskeletal dynamics

Background:

  • Cells dynamically interact with their mechanical environment, sensing and generating forces.
  • Understanding forces generated by non-adherent cells is crucial for cell biology.

Purpose of the Study:

  • To quantitatively measure forces and elastic properties of suspended 3T3 fibroblasts.
  • To investigate the role of non-muscle myosin II and microtubules in cellular force generation.

Main Methods:

  • Utilized a dual optical trap to suspend fibronectin-coated beads with 3T3 fibroblasts.
  • Measured cell stiffness and force fluctuations with high bandwidth.
  • Employed pharmacological inhibitors (blebbistatin, nocodazole) and serum starvation.

Main Results:

  • Cell stiffness significantly decreased with myosin inhibition and serum starvation.
  • Microtubule depolymerization did not affect cell stiffness.
  • Cortical forces generated by non-muscle myosin II deform cells (0.1–10 Hz); amplitudes reduced by blebbistatin and serum starvation.
  • Force fluctuations exhibited a spectrum characteristic of an elastic network with random stresses.

Conclusions:

  • Non-muscle myosin II is a primary determinant of cellular stiffness and force generation in suspended fibroblasts.
  • Microtubules play a minimal role in these specific mechanical properties.
  • Cellular force generation involves an elastic network activated by random stresses.