Distribution of Red Blood Cells Deformability: Study on Density-Separated Cell Subpopulations

Aging-related processes lead to significant metabolic and structural changes in red blood cells (RBCs) and, as a result, to heterogeneity in cell populations. Using the Percoll linear density gradient, separating the RBC population and obtaining fractions enriched with cells of different ages is possible. Previously, cells from the “light” fraction were characterized by increased deformability. However, the distribution of RBC deformability in subpopulations possessing a different density has not been studied. In this study, we measured the deformability of RBCs from cell fractions characterized by different densities. RBC deformability was determined using a computerized cell flow-properties analyzer, which provides the deformability distribution in a population of 10,000–15,000 cells. Our results demonstrate a strong correlation between the cytosol hemoglobin concentration and the cell deformation indexes. In addition, we show that the “lightest” fraction of RBCs contains the lowest number of deformable and the highest number of highly deformable cells. In contrast, the “dense” fraction is enriched with undeformable RBCs, with a minimal presence of highly deformable cells. In summary, we have shown that RBC fractions depleted or enriched with undeformable cells can be obtained by using a density gradient. However, these fractions are not homogeneous in their deformability properties.