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Direct methods for measuring microbial populations in a culture are essential tools in microbiology, providing quantitative data for various applications. Among these, microscopic counts, plate counts, and serial dilution are widely used techniques, each with unique principles and applications.Microscopic CountsMicroscopic counting involves the use of a Petroff-Hausser chamber, a specialized microscope slide with a grid and defined depth. By observing a liquid culture under a microscope,...
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Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding
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Macromolecular crowding limits growth under pressure.

Baptiste Alric1, Cécile Formosa-Dague2, Etienne Dague3

  • 1MILE team, CNRS, UPR8001, LAAS-CNRS, 7 Avenue du Colonel Roche, F-31400 Toulouse, France.

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|May 8, 2023
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Summary
This summary is machine-generated.

Cells under pressure from confined growth experience reduced growth rates. This study reveals increased macromolecular crowding as the cause, linking physical properties to cell behavior.

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Area of Science:

  • Biophysics
  • Cellular mechanics
  • Microbiology

Background:

  • Cells confined in space experience mechanical stress, known as growth-induced pressure (GIP).
  • GIP is implicated in various biological processes, including cancer and infections, but its origins and molecular effects are not fully understood.
  • Understanding GIP is crucial for comprehending cellular responses to environmental constraints.

Purpose of the Study:

  • To investigate the physical mechanisms underlying growth-induced pressure in confined cells.
  • To determine the relationship between mechanical stress and cellular properties.
  • To develop a predictive model for how GIP affects cell growth.

Main Methods:

  • Utilized microfluidic confinement of yeast (Saccharomyces cerevisiae).
  • Employed genetically encoded multimeric nanoparticles (GEMs) for rheological measurements.
  • Developed and calibrated a biophysical model to link macromolecular crowding to cell growth.

Main Results:

  • Demonstrated that growth-induced pressure correlates with increased macromolecular crowding within cells.
  • Showed that elevated macromolecular crowding hinders protein expression, leading to reduced cell growth.
  • Validated a model predicting growth rate reduction due to increased macromolecular crowding.

Conclusions:

  • Macromolecular crowding is a key physical property mediating the effects of growth-induced pressure on cell growth.
  • The observed feedback mechanism is independent of specific signaling pathways, suggesting a fundamental biophysical principle.
  • This mechanism of biomechanical feedback may be conserved across all life forms for environmental sensing.