Difference in Geometry of Young and Old Human Erythrocytes Explained by a Filtering Mechanism (original) (raw)
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Blood, 1992
The rheologic properties of senescent erythrocytes have been examined using two models of red blood cell (RBC) aging. In the rabbit, aged erythrocytes were isolated after biotinylation, in vivo aging, and subsequent recovery on an avidin support. Aged RBCs from the mouse were obtained using the Ganzoni hypertransfusion model that suppresses erythropoiesis for prolonged periods of time allowing preexisting cells to age in vivo. In both cases, the aged erythrocytes were found by ektacytometry to have decreased deformability due to diminished surface area and cellular dehydration. The aged rabbit erythrocytes were further characterized by micropipette methods that documented an average surface area decrease of 10.5% and a volume decrease of 8.4% for the cells that were 50 days old. Because both the surface area and volume decreased with cell age, there was little change in surface-to-volume ratio (sphericity) during aging. The aged cells were found to have normal membrane elasticity. I...
The development of a filtration system for evaluating flow characteristics of erythrocytes
Microvascular Research, 1987
A complete description of the pathophysiology of sickle cell disease requires a physiologically meaningful measurement of red cell deformability. We have designed and built a system which allows one to determine filtration characteristics of erythrocytes. A dilute red cell suspension is forced through a 3.0~pm polycarbonate Nuclepore membrane with a constant positive pressure of 20 mm Hg. Under these conditions blockage of the pores in the polycarbonate membrane is insignificant and flow is linear. We use the relative number of cells filtered through the membrane as a means of approximating the mean deformability of cells in the suspension. Using this system we have compared erythrocytes from various mammals and shown that our technique is sensitive in detecting not only differences in cell deformabilities between mammalian species but also changes in cell deformability of human red cells due to exchange transfusion and application of drugs. There was a positive correlation between cell filtrabihty and percentage cell recovery (coefficient of correlation, 0.65) and a negative correlation between cell size and filtrability (coefficient of correlation,-0.61). The filtrabilities of normal volunteers and sickle cell disease patients were found to be 71.8 ? 6.6 and 53.6 _' 5.0%, respectively. This system is sensitive and reliable, and should be useful in evaluating both the contribution of &ability to the viability of red cells in vivo and potential therapeutic agents for sickle cell disease.
2006
The use of microfabrication technology in the study of biological systems continues to grow rapidly in both prevalence and ascendancy. Customised microdevices that provide superior results than traditional macroscopic methods can be designed in order to investigate specific cell types and cellular processes. This study showed the benefit of this approach in precisely characterising the progressive losses of surface area and haemoglobin (Hb) content by the human red blood cell (RBC), from newborn reticulocyte to senescent erythrocyte. The high-throughput, multiparametric measurements made on individual cells with a specialised microdevice enabled, for the first time, delineation and quantification of the losses that occur during the two stages of the human RBC lifespan. Data acquired on tens of thousands of red cells showed that nearly as much membrane area is lost during the 1-2 d of reticulocyte maturation (c. 10-14%) as in the subsequent 4 months of erythrocyte ageing (c. 16-17%). The total decrease in Hb over the red cell lifespan is also estimated (c. 15%) and a model describing the complete timecourse of diminishing mean RBC area and Hb is proposed. The relationship between the losses of Hb and area, and their possible influence on red cell lifespan, are discussed.
Changes in the properties of normal human red blood cells during in vivo aging
American Journal of Hematology, 2013
The changes in red blood cells (RBC) as they age and the mechanisms for their eventual removal have been of interest for many years. Proposed age-related changes include dehydration with increased density and decreased size, increased membrane IgG, loss of membrane phospholipid asymmetry, and decreased activity of KCl cotransport. The biotin RBC label allows unambiguous identification of older cells and exploration of their properties as they age. Autologous normal human RBC were labeled ex vivo and, after reinfusion, compared with unlabeled RBC throughout their lifespan. RBC density increased with age, with most of the change in the first weeks. Near the end of their lifespan, RBC had increased surface IgG. However, there was no evidence for elevated external phosphatidylserine (PS) even though older RBC had significantly lower activity of aminophospholipid translocase (APLT). KCl cotransport activity persisted well past the reticulocyte stage, but eventually decreased as the RBC became older. These studies place limitations on the use of density fractionation for the study of older human RBC, and do not support loss of phospholipid asymmetry as a mechanism for human RBC senescence. However, increased levels of IgG were associated with older RBC, and may contribute to their removal from the circulation. Am.
In Vivo Volume and Hemoglobin Dynamics of Human Red Blood Cells
2014
Human red blood cells (RBCs) lose ~30% of their volume and ~20% of their hemoglobin (Hb) content during their ~100-day lifespan in the bloodstream. These observations are well-documented, but the mechanisms for these volume and hemoglobin loss events are not clear. RBCs shed hemoglobin-containing vesicles during their life in the circulation, and this process is thought to dominate the changes in the RBC physical characteristics occurring during maturation. We combine theory with single-cell measurements to investigate the impact of vesiculation on the reduction in volume, Hb mass, and membrane. We show that vesicle shedding alone is sufficient to explain membrane losses but not volume or Hb losses. We use dry mass measurements of human RBCs to validate the models and to propose that additional unknown mechanisms control volume and Hb reduction and are responsible for ~90% of the observed reduction. RBC population characteristics are used in the clinic to monitor and diagnose a wide range of conditions including malnutrition, inflammation, and cancer. Quantitative characterization of cellular maturation processes may help in the early detection of clinical conditions where maturation patterns are altered.
Biophysical journal, 2009
Erythrocytes (red blood cells) play an essential role in the respiratory functions of vertebrates, carrying oxygen from lungs to tissues and CO 2 from tissues to lungs. They are mechanically very soft, enabling circulation through small capillaries. The small thermally induced displacements of the membrane provide an important tool in the investigation of the mechanics of the cell membrane. However, despite numerous studies, uncertainties in the interpretation of the data, and in the values derived for the main parameters of cell mechanics, have rendered past conclusions from the fluctuation approach somewhat controversial. Here we revisit the experimental method and theoretical analysis of fluctuations, to adapt them to the case of cell contour fluctuations, which are readily observable experimentally. This enables direct measurements of membrane tension, of bending modulus, and of the viscosity of the cell cytoplasm. Of the various factors that influence the mechanical properties of the cell, we focus here on: 1), the level of oxygenation, as monitored by Raman spectrometry; 2), cell shape; and 3), the concentration of hemoglobin. The results show that, contrary to previous reports, there is no significant difference in cell tension and bending modulus between oxygenated and deoxygenated states, in line with the softness requirement for optimal circulatory flow in both states. On the other hand, tension and bending moduli of discocyte-and spherocyteshaped cells differ markedly, in both the oxygenated and deoxygenated states. The tension in spherocytes is much higher, consistent with recent theoretical models that describe the transitions between red blood cell shapes as a function of membrane tension. Cell cytoplasmic viscosity is strongly influenced by the hydration state. The implications of these results to circulatory flow dynamics in physiological and pathological conditions are discussed.
The Journal of Physiology, 1978
The observation that human red blood cells do not shrink in hypertonic media as much as expected for ideal osmometers has previously been explained in terms of either a marked increase in the osmotic coefficient of the cell contents or an increase in the chloride content of the cells. 2. Changes in suspension pH and haematocrit have been observed when the concentration of the unbuffered NaCl medium was doubled. The small increases in external pH, and the size of the volume decreases, are inconsistent with variations in the Cl content as a significant factor in the non-ideal osmotic responses. 3. Membrane potentials of red cells in buffered media were followed using the fluorescent dye, diS-C3-(5). On shrinking at pH 7 4, the cells hyperpolarized ca. 5 mV as predicted if changes in the osmotic coefficient rather than in Cl content explained the osmotic behaviour. 4. Regarding haemoglobin in concentrated solution as a solute with high osmotic coefficient is formally correct but is little help in understanding the properties of the solution. We have found it useful to consider separately haemoglobin and the rest of the contents of the cell. The haemoglobin then supports part of the total hydrostatic pressure on the cell leaving the crystalloid solution to experience a reduced fluid pressure. In greatly shrunken cells the contents act like a gel with the matrix of haemoglobin under compression and the fluid which fills the spaces within the matrix under tension. Since there are neither hydrostatic (Rand & Burton, 1964) nor osmotic (Williams, Fordham, Hollander & Welt, 1959) pressure gradients across the human red cell membrane, there are only two possible types of explanation for the observed volume
Correlation between local cell membrane displacements and filterability of human red blood cells
FEBS Letters, 1992
Local mechanical fluctuations of the cell membrane of human erythrocytes were shown to involve MgATP-and Mg"+-driven fast membrane displacements. We propose that these local bending deformations of the cell membrane arc important for cell passage through capillaries. In order to verify this hypothesis, we examined cell membrane fluctuations and filterability ol'crythrocy~cs over a wide mngeofmcdium osmolaliries( 180-675 mosmol/kg H20). The results indicate the existence of a positive correlalion between the amplitude of local cell membrane displacements and cell filterability. WC suggest that the occurrence of metabolically driven membrane displacements on the side surface of the red blood cell diminishes its bending stiffness and enables it to fold more efficiently upon entrance into blood capillaries. Thus, local cell membrane displacements seem to play an important role in microcirculation.
Osmotic fragility model for red cell populations
Biophysical Journal, 1988
A model that predicts the osmotic fragility curve of a red cell population is developed by relating the critical osmotic pressure to the size distribution of the cells, determined by resistive pulse spectroscopy. Two of the parameters involved, namely the normalized osmotic volume correction, B, and the swelling index, k, are previously determined from the experimental average properties of the population. From these values the critical volume of the cell is obtained, and is shown to be 6-12% larger than the first spherical volume, obtained from an independent experiment. A new parameter, n, a measure of the surface area distribution of the cells, is incorporated through a simple function that relates the critical volume to the size of the cells, and is theoretically shown to be linked to parameters k and B. The model is used to fit and interpret fragility data obtained in this laboratory for normal and sickle cell samples. From the values of n obtained for normal samples, the model predicts an essentially constant surface-to-volume ratio within an individual's cell population. For sickle cell samples, instead, the value of index n is negative, thereby supporting an increase in excess surface area as cell size decreases. Both findings are in agreement with direct observations reported in the literature. It is concluded that this set of parameters may be used to develop an index classification of blood disorders.