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Related Experiment Videos

Endothelial cells reestablish functional integrity after reversible permeabilization.

D F Fennell1, R E Whatley, T M McIntyre

  • 1Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah, Salt Lake City 84112.

Arteriosclerosis and Thrombosis : a Journal of Vascular Biology
|January 1, 1991
PubMed
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Endothelial cells (ECs) can be permeabilized using glass beads, allowing intracellular molecule introduction without cell removal. These transiently porated ECs regain function, aiding studies of endothelial repair and regulatory mechanisms.

Area of Science:

  • Cell Biology
  • Biophysics

Background:

  • Permeabilization is crucial for manipulating intracellular mechanisms in cell biology.
  • Introducing probes and molecules into the cell cytoplasm requires effective permeabilization techniques.

Purpose of the Study:

  • To investigate glass bead-induced, non-specific permeabilization of endothelial cells (ECs).
  • To determine the conditions for reestablishing plasma membrane integrity in porated ECs.
  • To assess the functional recovery of permeabilized ECs.

Main Methods:

  • Incubation of human and bovine ECs with glass beads in monolayer culture.
  • Introduction of macromolecules (dextrans, immunoglobulins) and small molecules (Lucifer Yellow).
  • Assessment of membrane resealing based on time, temperature, and extracellular calcium.

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Main Results:

  • Glass bead incubation caused transient, non-specific plasma membrane permeabilization in ECs.
  • Macromolecules and small charged molecules were successfully introduced into the cytoplasm.
  • Plasma membrane integrity was reestablished, dependent on time, temperature, and calcium.
  • Permeabilized ECs regained functional characteristics, including monolayer formation and molecule extrusion.

Conclusions:

  • Transient EC permeabilization with glass beads allows intracellular loading while maintaining monolayer culture.
  • Permeabilized ECs serve as a model for studying endothelial plasma membrane repair mechanisms.
  • This method facilitates research on intracellular regulatory mechanisms in situ-like conditions.