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Measuring Deformability and Red Cell Heterogeneity in Blood by Ektacytometry
Published on: January 12, 2018
Biophysical and Biochemical Markers of Red Blood Cell Fragility
Ariel Orbach1, Orly Zelig2, Saul Yedgar1
1Department of Biochemistry, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Background:
Red blood cells (RBCs) undergo a natural aging process occurring in the blood circulation throughout the RBC lifespan or during routine cold storage in the blood bank. The aging of RBCs is associated with the elevation of mechanical fragility (MF) or osmotic fragility (OF) of RBCs, which can lead to cell lysis. The present study was undertaken to identify RBC properties that characterize their susceptibility to destruction under osmotic/mechanical stress.
Methods:
RBCs were isolated from freshly donated blood or units of packed RBCs (PRBCs) and suspended in albumin-supplemented phosphate-buffered saline (PBS). In addition, PRBCs were separated by filtration through a microsphere column into two fractions: enriched with rigid (R-fraction) and deformable (D-fraction) cells. The RBCs were subjected to determination of deformability, MF and OF, moreover, the level of cell surface phosphatidylserine (PS) and the stomatin level in isolated RBC membranes were measured.
Results:
In the RBC population, the cells that were susceptible to mechanical and osmotic stress were characterized by low deformability and increased level of surface PS. The OF/MF was higher in the R-fraction than in the D-fraction. Stomatin was depleted in destroyed cells and in the R-fraction.
Conclusion:
RBC deformability, the levels of surface PS, and membrane stomatin can be used as markers of RBC fragility.
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