Bregt Roerdinkholder-Stoelwinder

Radboud University Medical Centre (Radboudumc), Nymegen, Gelderland, Netherlands

Are you Bregt Roerdinkholder-Stoelwinder?

Claim your profile

Publications (12)54.18 Total impact

    [Show abstract] [Hide abstract]
    ABSTRACT: Summary A recently described separation technique consisting of a combination of counterflow centrifugation and subsequent density (Percoll) scparation was tested for its ability to enrich red cell populations with young cells in comparison to either separation technique alone. The relative age of every fraction was determined by HbAlc measurements, resulting in the lowest HbAlc for the combination method. Conventional reticulocyte counting and floweytometric counting with thiazole orange indicated that in the youngest fractions the combination method showed the highest reticulocyte counts. There was a good correlation between manual and flowcytometric counting results. Radio-iron studies showed a two-fold enrichment with young cells in the fraction with the lowest HbAIc acquired by the combination technique in comparison to the other two methods. Cytometric measurements showed that the fractions with the lowest HbAlc were the ones with the highest MCV and MCH and the lowest MCHC. Besides loss of their RNA-material, young cells already seem to loose water and haemoglobin like older cells, resulting in a decrease of MCV and MCH and in increase in MCHC. It is concluded that combining counterflow centrifugation with subsequent density fractionation results in superior enrichment with young cells in comparison to the results of each method alone.
    No preview · Article · Jun 2008 · Clinical & Laboratory Haematology
  • [Show abstract] [Hide abstract]
    ABSTRACT: Previous studies demonstrated that 20% of haemoglobin is lost from circulating erythrocytes during their total lifespan by vesiculation. To study whether removal molecules other than membrane-bound haemoglobin were present in erythrocyte-derived vesicles, flow cytometry and immunoblot analysis were employed to examine the presence of phosphatidylserine (PS) and IgG, and senescent cell antigens respectively. It was demonstrated that 67% of glycophorin A-positive vesicles exposed PS, and that half of these vesicles also contained IgG. Immunoblot analysis revealed the presence of a breakdown product of band 3 that reacted with antibodies directed against senescent erythrocyte antigen-associated band 3 sequences. In contrast, only the oldest erythrocytes contained senescent cell antigens and IgG, and only 0.1% of erythrocytes, of all ages, exposed PS. It was concluded that vesiculation constitutes a mechanism for the removal of erythrocyte membrane patches containing removal molecules, thereby postponing the untimely elimination of otherwise healthy erythrocytes. Consequently, these same removal molecules mediate the rapid removal of erythrocyte-derived vesicles from the circulation.
    No preview · Article · Jun 2008 · British Journal of Haematology
  • Marleen Luten · Bregt Roerdinkholder-Stoelwinder · Nicolaas P.M. Schaap · Willem J de Grip · Harry J Bos · Giel J.C.G.M. Bosman
    [Show abstract] [Hide abstract]
    ABSTRACT: The use of fresh red blood cells (RBCs) is recommended for critically ill patients and patients undergoing surgery, although there is no conclusive evidence that this is beneficial. In this follow-up study, the short-term and the long-term recovery of irradiated, leukoreduced RBCs transfused after either a short storage (SS) or a long storage (LS) period were compared. By consecutive transfusion of RBCs with a SS and LS period, a direct comparison of their survival within the same patient was possible. Ten transfusion-requiring patients each received a SS RCCs (stored 0-10 days) and a LS RCCs (stored 25-35 days) consecutively. Short-term and long-term survival of the transfused RBCs was followed by flow cytometry using natural differences in RBC antigens between donors and patients. Posttransfusion recovery (PTR) was measured at several time points after transfusion. The mean 24-hour PTR of SS RBCs is 86.4 +/- 17.8 percent and that of LS RBCs 73.5 +/- 13.7 percent. After the first 24 hours, the mean times to reach a PTR of 50 percent of the 24-hour PTR (T50) and mean potential life spans (mPLs) of the surviving SS and LS RBCs (41 and 116 days and 41 and 114 days, respectively) do not differ. The mean 24-hour PTR of both SS and LS RBCs complies with the guidelines, even in a compromised patient population. The 24-hour PTR of SS RBCs, however, is significantly higher than that of LS RBCs. The remaining population of SS and LS RBCs has a nearly identical long-term survival. Therefore, depletion of the removal-prone RBCs before transfusion may be an efficient approach for product improvement.
    No preview · Article · Jun 2008 · Transfusion
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: During storage of red cells (RBCs) for transfusion, RBCs undergo a number of biochemical and morphologic changes. To be able to identify the mechanisms underlying these storage lesions, a proteomic analysis of the membranes of RBCs and their vesicles was performed during various periods of storage in blood bank conditions. RBCs and vesicles were isolated from RBCs after various storage periods. The proteins of RBC membranes and vesicles were separated by gel electrophoresis and identified by a semiquantitative proteomic analysis. Our findings confirm previous data, such as a storage-associated increase in hemoglobin binding to the membrane and aggregation and degradation of the integral membrane protein band 3, suggesting a remodeling of the RBC membrane during storage. Our data also show storage-dependent changes in the membrane association of proteasome and chaperone proteins, metabolic enzymes, small G proteins, and signal transduction proteins. Vesicles display similar changes in their protein composition during storage. The results of this analysis indicate that the storage-related changes in the RBC membrane are the results of disturbance and/or acceleration of physiologic processes such as cellular aging, including vesicle formation. The latter may serve to remove damaged membrane patches that would otherwise lead to accelerated RBC removal. These data provide a framework for future studies toward the development of better storage conditions and a reduction of the side effects of RBC transfusion.
    Full-text · Article · Jun 2008 · Transfusion
  • [Show abstract] [Hide abstract]
    ABSTRACT: Therapeutic phlebotomy is the preferred treatment for iron overload associated with hemochromatosis. In the Netherlands, red blood cell concentrates (RCCs) from hemochromatosis patients are not used for transfusion purposes. In this study, their storage performance was compared with that of control donors as a first step in the evaluation of their potential usefulness for transfusion. RCCs were obtained from hemochromatosis patients and regular donors, either by apheresis or by whole-blood collection, and stored up to 50 days under routine Dutch blood bank conditions. Weekly samples were taken for determination of hematologic, biophysical, and biochemical variables. Most variables displayed the same storage-related changes in RCCs originating from hemochromatosis patients as in those from regular donors. In all RCCs, hemolysis remained well below the guideline limit of 0.8 percent for up to 6 weeks of storage, and the glucose concentration remained above the required 5 mmol per L up to 5 weeks of storage. After 4 weeks of storage, the mean ATP level remained above the required limit of 75 percent of the starting value in all RCCs as well. The major difference was a larger mean cell volume in hereditary hemochromatosis RBCs up to 50 days of storage. RCCs from hemochromatosis patients comply with the in vitro quality requirements for transfusion. This paves the way for the final step, namely, the establishment of the 24-hour RBC posttransfusion recovery.
    No preview · Article · Apr 2008 · Transfusion
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Previous studies have shown that during the lifespan of red blood cells (RBCs) 20% of hemoglobin is lost by shedding of hemoglobin-containing vesicles. However, the fate of these vesicles is unknown. To study this fate we used a rat model, after having established that rat RBCs lose hemoglobin in the same way as human RBCs, and that RBC-derived vesicles are preferentially labeled by Na2(51) CrO4. Such labeled vesicles were injected into recipient rats. Within 5 minutes, 80% of the radioactivity was cleared from the circulation with a concomitant uptake by the liver of 55% of the injected dose. After 30 minutes, Kupffer cells contained considerable amounts of hemoglobin and were shown to be responsible for 92% of the liver uptake. Vesicle clearance from the blood as well as liver uptake were significantly inhibited by preinjection of the scavenger-receptor ligands polyinosinic acid and phosphatidylserine. We conclude that in rats Kupffer cells rapidly remove RBC-derived vesicles from the circulation, mainly by scavenger receptors. The same mechanism is likely to be responsible for the elimination of human RBC vesicles, thereby constituting an important pathway for the breakdown of RBCs in humans.
    Full-text · Article · Apr 2005 · Blood
  • Source
    M Luten · B Roerdinkholder-Stoelwinder · H J Bost · G J C G M Bosman
    [Show abstract] [Hide abstract]
    ABSTRACT: During the last 90 years many developments have taken place in the world of blood transfusion. Several anticoagulants and storage solutions have been developed. Also the blood processing has undergone many changes. At the moment, in The Netherlands, red blood cell (RBC) concentrates (prepared from a whole blood donation and leukocyte-depleted by filtration) are stored for a maximum of 35 days at 4 degrees C in saline adenine glucose mannitol (SAGM). Most relevant studies show that approximately 20% of the RBCs is lost in the first 24 hr after transfusion. Even more remarkable is that the average life span is 94 days after a storage period of 42-49 days. Such observations create the need for a parameter to measure the biological age of RBCs as a possible predictor of the fate of RBCs after transfusion. The binding of IgG to RBCs can lead to recognition and subsequent phagocytosis by macrophages. This occurs during the final stages of the RBC life span in vivo. We determined the quantity of cell-bound IgG during storage, and found considerable variation between RBCs, but no significant storage-related change in the quantity of cell-bound IgG. The significance of this finding for predicting the survival of transfused RBCs in vivo remains to be established. Hereto we developed a flow cytometric determination with a sensitivity of 0.1% for the measurement of survival in vivo based on antigenic differences. This technique has various advantages compared with the 'classical' 51Cr survival method.
    Full-text · Article · Apr 2004 · Cellular and molecular biology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Previous studies have shown that approximately 20% of hemoglobin is lost from circulating red blood cells (RBCs), mainly during the second half of the cells' life span. Because hemoglobin-containing vesicles are known to circulate in plasma, these vesicles were isolated. Flow cytometry studies showed that most RBC-derived vesicles contain hemoglobin with all hemoglobin components present. The hemoglobin composition of the vesicles resembled that of old RBCs. RBC cohort studies using isotope-labeled glycine have been described, which showed a continuous presence of this label in hemoglobin degradation products. The label concentration of these products increased during the second half of the RBC life span, accompanied by a decrease within the RBC. It is concluded that the hemoglobin loss from circulating RBCs of all ages can be explained by shedding hemoglobin-containing vesicles. This loss occurs predominantly in older RBCs. Apparently the spleen facilitates this process since asplenia vesicle retention within RBCs of all ages has been described, accompanied by an increase in the percentage of total HbA(1). The present study shows that in old RBCs of asplenic individuals, the decrease of hemoglobin content per cell such as seen in old RBCs of control individuals is absent due to an increase in the absolute amount of HbA(1c) and HbA(1e2). It is concluded that hemoglobin-containing vesicles within old RBCs are "pitted" by the spleen.
    Full-text · Article · Feb 2003 · Blood
  • F. L. A. Willekens · F H Bosch · B Roerdinkholder-Stoelwinder · Y. A. M. Groenen-Döpp · J M Werre
    [Show abstract] [Hide abstract]
    ABSTRACT: Previous studies have shown that a considerable amount of haemoglobin is lost from the intact red cell during its lifespan. The aim of this study was to determine the relative contribution of all the haemoglobin components to this process. Therefore, the relative amount of haemoglobins A0, A2, F and the glycated haemoglobins were determined in 24 fractions of different cell age. These fractions were obtained by the combination of counterflow and density centrifugation. When the absolute amount of all haemoglobin components were calculated using the MCH-values of each fraction, it appeared that the mean red cell loss of haemoglobins A0, A2, F, an unknown X and "rest" comprised, respectively, 440, 23, 1, 4 and 1 amol per cell, while the mean gain of the glycated haemoglobins was 84 amol per cell. This resulted in a net loss of 385 amol of haemoglobin per cell. One of the glycated haemoglobins (HbA1e2) turned out to be the product of further carbamylation. It was concluded that in the first half of the red cell lifespan HbA0 and HbA2 decreased by glycation and carbamylation and that in the second half some of the HbA0 and HbA2 but also some of the glycated and carbamylated haemoglobin components leave the red cell. The total loss amounted to about 20%.
    No preview · Article · May 1997 · European Journal Of Haematology
  • [Show abstract] [Hide abstract]
    ABSTRACT: Introduction: Loss of lipid asymmetry in the membranes of nucleated cells by exposure of phosphatidylserine is thought to be a crucial part of the process of apoptosis of those cells. Red cells also lose the phospholipid asymmetry of their plasma membrane during incubation in vitro. Purpose: Is lipid asymmetry also lost in RBC's during their life span in vivo? Methods: Red cells were separated in five fractions of different cell age (1-V) by a combination of percoll gradient- and counter- flow centrifugation. FITC-labelled annexin V was used to detect phosphatidylserine in the outer layer of the red cell membranes by means of a flow cytometric technique (FACS). As markers of RBC age the percentage of HbA1c was used. Results: In fractions I to V: 1) HbA1c increased from 3.73 to 6.71; 2) The percentage of annexin V positive cells did not increase (see Table). Mean of annexin-V positive RBC's in whole blood and in fractions of different cell age of normal individuals (n=6) RBC's HbA1c (% ± SD) Annexin positive cells (% ± SD) Whole blood 5.20 ± 0.35 0.11 ± 0.04 Fraction I 3.73 ± 0.54 0.26 ± 0.17 Fraction II 4.47 ± 0.39 0.14 ± 0.04 Fraction III 5.28 ± 0.54 0.15 ± 0.08 Fraction IV 6.21 ± 0.64 0.12 ± 0.05 Fraction V 6.71 ± 0.59 0.12 ± 0.03 Controls showed more than 50% positive cells. Conclusion: The phosphatidylserine concentration of the outer layer of RBC's was not increased in fractions enriched with old cells indicating that in this study there was no evidence for loss of lipid asymmetry in circulating old red cells as cause of cell death. This is in contrast to the change in the membranes of RBC's in vitro and of certain nucleated cells during apoptosis.
    No preview · Article · Jan 1997
  • J.M. Werre · F.L.A. Willekens · H.J. Bos · B. Roerdinkholder-Stoelwinder · Y.A.M. Groenen-Dopp · G.C. van der Plas · A.J. Verkley

    No preview · Article · Jan 1996 · The Netherlands Journal of Medicine
  • F H Bosch · J M Werre · L Schipper · B Roerdinkholder-Stoelwinder · T. H. Huls · F.L.A. Willekens · G Wichers · M R Halie
    [Show abstract] [Hide abstract]
    ABSTRACT: Red blood cell (RBC) deformability was determined with an ektacytometer in fractions separated on the basis of differences in cell volume or density. Deformability was measured with ektacytometry (rpm-scan and osmo-scan). We studied three groups of RBC fractions:1. By counterflow centrifugation we obtained fractions of different cell age which showed a slight decrease in mean corpuscular haemoglobin concentration (MCHC) and an increase in surface-to-volume (S/V) ratio in fractions with older cells. 2. By Percoll fractionation fractions were obtained which showed a pronounced increase in (MCHC) but no change in S/V ratio. 3. By a combination of both fractionation techniques, fractions were obtained which showed an increased MCHC and an increase in S/V ratio. Deformability in group 1,2 and 3 showed respectively no change, a moderate decrease and a pronounced decrease in fractions of older cells. A decline in deformability occurs during the aging process of the red blood cell. This decline in deformability in old red cells is greater than originally thought. This decline is the result of an increase in haemoglobin concentration and a second factor, probably a decrease in membrane elasticity.
    No preview · Article · Feb 1994 · European Journal Of Haematology
  • Source
    F H Bosch · J M Werre · B Roerdinkholder-Stoelwinder · T H Huls · F.L.A. Willekens · M R Halie
    [Show abstract] [Hide abstract]
    ABSTRACT: Red blood cell (RBC) fractions were studied after separation of whole blood by means of counterflow centrifugation, Percoll column (Pharmacia, Uppsala, Sweden), and a combination of both separation techniques. Mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH), and hemoglobin A1c (HbA1c) were measured in each fraction. From the results it was obvious that the combination of both techniques was the best separation technique of these three. MCV had a good correlation with cell age as measured with HbA1c concentration gradient; MCH and MCHC less so. MCV and MCH decreased in parallel to an increase in HbA1c. MCHC increased with increasing HbA1c. From these data it is concluded that there is a steadily ongoing loss of cellular hemoglobin and proportionally more cellular water during the life of the RBC.
    Preview · Article · Feb 1992 · Blood

Publication Stats

653 Citations
54.18 Total Impact Points


  • 2008
    • Radboud University Medical Centre (Radboudumc)
      • Department of Biochemistry
      Nymegen, Gelderland, Netherlands
  • 1992-2008
    • Rijnstate Hospital
      Arnheim, Gelderland, Netherlands
  • 1997
    • Red Cross
      Washington, Washington, D.C., United States