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Related Experiment Videos

Extracellular matrix- and cytoskeleton-dependent changes in cell shape and stiffness.

Kiran Bhadriraju1, Linda K Hansen

  • 1Biomedical Engineering Institute, MMC 609, 420 Delawere Street S. E., University of Minnesota, Minneapolis, Minnesota 55455, USA.

Experimental Cell Research
|July 20, 2002
PubMed
Summary

Hepatocyte cell stiffness increases with spreading, requiring normal actin and myosin function for both spreading and DNA synthesis. Disrupting actin or myosin impacts cell stiffness differently, highlighting their complex roles.

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Area of Science:

  • Cell biology
  • Biophysics
  • Hepatocyte research

Background:

  • Cell spreading influences key cellular functions like DNA synthesis and differentiation.
  • Cytoskeletal reorganization during cell spreading affects cell stiffness, particularly in motile cells.
  • Limited data exists on cytoskeletal dynamics and stiffness changes in nonmigratory epithelial cells.

Purpose of the Study:

  • To investigate the relationship between cell function, spreading, and stiffness in hepatocytes.
  • To understand how fibronectin density influences hepatocyte spreading and mechanical properties.
  • To elucidate the distinct roles of actin and myosin in hepatocyte spreading, stiffness, and DNA synthesis.

Main Methods:

  • Atomic force microscopy (AFM) was used to measure cell stiffness.

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  • Hepatocytes were cultured on fibronectin-coated surfaces with varying densities (1 and 1000 ng/cm(2)).
  • Pharmacological agents were used to disrupt actin and myosin function.

    • Main Results:

    • Cell stiffness increased with spreading on high fibronectin density but remained low when cells were rounded on low fibronectin density.
    • Disrupting actin or myosin inhibited cell spreading and DNA synthesis.
    • Disrupting f-actin assembly decreased both stiffness and spreading; inhibiting myosin light chain kinase decreased spreading but increased stiffness.

    Conclusions:

    • Normal actin and myosin function is essential for hepatocyte spreading and DNA synthesis.
    • Cell stiffness is directly related to cell spreading in hepatocytes.
    • Disruption of actin and myosin pathways has differential effects on hepatocyte stiffness.