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Systematic Analysis of In Vitro Cell Rolling Using a Multi-well Plate Microfluidic System
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Dimethylsulfoxide exposure modulates HL-60 cell rolling interactions.

David J Gee1, L Kate Wright, Jonathan Zimmermann

  • 1Department of Mechanical Engineering, Rochester Institute of Technology, 76 Lomb Memorial Drive, Rochester, NY, USA. David_Gee@mail.RIT.edu

Bioscience Reports
|April 13, 2012
PubMed
Summary

Dimethyl sulfoxide (DMSO) treatment alters human leukaemic HL-60 cell rolling velocity, potentially impacting their use in studying leukocyte adhesion cascade dynamics.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Immunology

Background:

  • Human leukaemic HL-60 cells mimic leucocyte rolling dynamics in vitro.
  • These cells are used to study adhesion molecules like P-selectin and P-selectin glycoprotein ligand-1 (PSGL-1).
  • HL-60 cells differentiate into granulocytes upon exposure to dimethyl sulfoxide (DMSO).

Purpose of the Study:

  • To investigate the rolling behavior of undifferentiated and DMSO-induced HL-60 cells.
  • To assess the impact of DMSO-induced differentiation on HL-60 cell rolling dynamics.
  • To evaluate the suitability of differentiated HL-60 cells as a model for neutrophil trafficking.

Main Methods:

  • Utilized a parallel plate flow chamber with P-selectin-Fc chimaera.
  • Assayed rolling behavior of undifferentiated and DMSO-treated HL-60 cells (48, 72, 96 hours).
  • Flow cytometry was used to analyze PSGL-1 expression.

Main Results:

  • Undifferentiated cells showed increased rolling velocity with higher P-selectin concentration and shear stress.
  • Short-term DMSO treatment (up to 72h) increased rolling velocity.
  • Long-term DMSO treatment (96h) significantly decreased rolling velocity.
  • PSGL-1 expression remained unchanged despite DMSO treatment.

Conclusions:

  • DMSO-treated HL-60 cells exhibit altered rolling behavior compared to undifferentiated cells.
  • Cell surface remodeling during differentiation may affect rolling dynamics.
  • Differentiated HL-60 cells may not be ideal substitutes for neutrophils in advanced leukocyte adhesion cascade studies.