Transport of Na+ and HCO3- Out of Red Blood Cells Is Simultaneous with a Chloride Shift in Canine and Human Whole Blood Exposed to CO2-Rich Gas.

Background The SAME device (i-SEP, France) is an innovative filtration-based autotransfusion device able to salvage and wash both red blood cells and platelets. This study evaluated the device performances using human whole blood with the hypothesis that the device will be able to salvage platelets while achieving a erythrocyte yield of 80% and removal ratios of 90% for heparin and 80% for major plasma proteins without inducing signification activation of salvaged cells. Methods Thirty healthy human whole blood units (median volume, 478 ml) were diluted, heparinized, and processed by the device in two consecutive treatment cycles. Samples from the collection reservoir and the concentrated blood were analyzed. Complete blood count was performed to measure blood cell recovery rates. Flow cytometry evaluated the activation state and function of platelets and leukocytes. Heparin and plasma proteins were measured to assess washing performance. Results The global erythrocyte yield was 88.1% (84.1 to 91.1%; median [25th to 75th]) with posttreatment hematocrits of 48.9% (44.8 to 51.4%) and 51.4% (48.4 to 53.2%) for the first and second cycles, respectively. Ektacytometry did not show evidence of erythrocyte alteration. Platelet recovery was 36.8% (26.3 to 43.4%), with posttreatment counts of 88 × 109/l (73 to 101 × 109/l) and 115 × 109/l (95 to 135 × 109/l) for the first and second cycles, respectively. Recovered platelets showed a low basal P-selectin expression at 10.8% (8.1 to 15.2%) and a strong response to thrombin-activating peptide. Leukocyte yield was 93.0% (90.1 to 95.7%) with no activation or cell death. Global removal ratios were 98.3% (97.8 to 98.9%), 98.2% (96.9 to 98.8%), and 88.3% (86.6 to 90.7%) for heparin, albumin, and fibrinogen, respectively. The processing times were 4.4 min (4.2 to 4.6 min) and 4.4 min (4.2 to 4.7 min) for the first and second cycles, respectively. Conclusions This study demonstrated the performance of the SAME device. Platelets and red blood cells were salvaged without significant impact on cell integrity and function. In the meantime, leukocytes were not activated, and the washing quality of the device prevented reinfusion of high concentrations of heparin and plasma proteins. Editor’s Perspective What We Already Know about This Topic What This Article Tells Us That Is New

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A Simplified Method for the Determination of Omega-6/Omega-3 Ratios in Red Blood Cells from Human Whole Blood

10.35840/2633-8912/2417 ◽

2021 ◽

Vol 3 (1) ◽

Author(s):

Eberhart B Loye ◽

Wilson Annette S ◽

O'Keefe Stephen J D

Keyword(s):

Red Blood Cells ◽

Whole Blood ◽

Blood Cells ◽

Omega 3 ◽

Human Whole Blood ◽

Simplified Method ◽

Omega 6

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Carbon Isotopes Profiles of Human Whole Blood, Plasma, Red Blood Cells, Urine and Feces for Biological/Biomedical14C-Accelerator Mass Spectrometry Applications

Analytical Chemistry ◽

10.1021/ac103038s ◽

2011 ◽

Vol 83 (9) ◽

pp. 3312-3318 ◽

Cited By ~ 11

Author(s):

Seung-Hyun Kim ◽

Jennifer C. Chuang ◽

Peter B. Kelly ◽

Andrew J. Clifford

Keyword(s):

Mass Spectrometry ◽

Blood Plasma ◽

Red Blood Cells ◽

Carbon Isotopes ◽

Accelerator Mass Spectrometry ◽

Whole Blood ◽

Blood Cells ◽

Human Whole Blood

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Importance of L-selectin–dependent leukocyte–leukocyte interactions in human whole blood

Blood ◽

10.1182/blood.v95.9.2954.009k28_2954_2959 ◽

2000 ◽

Vol 95 (9) ◽

pp. 2954-2959 ◽

Cited By ~ 28

Author(s):

Debra J. Mitchell ◽

Pauline Li ◽

Paul H. Reinhardt ◽

Paul Kubes

Keyword(s):

Red Blood Cells ◽

Whole Blood ◽

Blood Cells ◽

Rhodamine 6G ◽

Model System ◽

Human Whole Blood ◽

Secondary Interactions ◽

First Time ◽

Rolling Leukocytes ◽

Deficient Mice

The objective of this study was to investigate whether leukocytes could be recruited by rolling leukocytes in a human whole blood model system. In all experiments, either neutrophils, whole blood, or diluted blood was perfused over immobilized E-selectin. With isolated neutrophils (2 × 105/mL), the free-flowing neutrophils were captured by attached neutrophils to form secondary interactions that resulted in lines of rolling leukocytes. These secondary tethers accounted for 50% to 60% of all interactions and were eliminated by an L-selectin antibody, which also eliminated the lines of rolling leukocytes. Perfusion of whole blood or diluted blood revealed no lines of rolling leukocytes. The addition of red blood cells to isolated neutrophils either in a 1000:1 or a 10:1 ratio also inhibited lines of rolling leukocytes. Leukocytes were fluorescently labeled with rhodamine-6G so that leukocyte–leukocyte interactions could be studied in whole blood. A small number of secondary tethers (less than 20%) occurred and could be reduced by more than 80% with an L-selectin antibody. However, the overall impact on leukocyte recruitment was negligible. Similar experiments were performed using murine whole blood or isolated murine leukocytes. In the absence of red blood cells, murine leukocytes also formed lines of rolling leukocytes on E-selectin, and secondary tethers accounted for 50% of total interactions. However, when murine blood (diluted 1:5 with buffer) was perfused over E-selectin, secondary tethers accounted for only 13% of total interactions. These interactions were completely absent when blood was used from L-selectin–deficient mice. These data demonstrate for the first time that the importance of L-selectin–dependent leukocyte–leukocyte interactions is greatly reduced in whole blood and does not enhance overall recruitment of leukocytes in this physiologic milieu.

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Determination of irinotecan (CPT-11) and SN-38 in human whole blood and red blood cells by liquid chromatography with fluorescence detection

Journal of Chromatography B ◽

10.1016/s1570-0232(03)00574-9 ◽

2003 ◽

Vol 795 (2) ◽

pp. 383-388 ◽

Cited By ~ 27

Author(s):

Floris A. de Jong ◽

Ron H.J. Mathijssen ◽

Peter de Bruijn ◽

Walter J. Loos ◽

Jaap Verweij ◽

...

Keyword(s):

Liquid Chromatography ◽

Red Blood Cells ◽

Fluorescence Detection ◽

Whole Blood ◽

Blood Cells ◽

Human Whole Blood

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An Evaluation of Morphological Changes and Deformability of Suspended Red Blood Cells Prepared Using Whole Blood with Different Hemoglobin Levels of Tibetans

Transfusion Medicine and Hemotherapy ◽

10.1159/000513319 ◽

2021 ◽

pp. 1-10

Author(s):

Rui Zhong ◽

Dingding Han ◽

Xiaodong Wu ◽

Hong Wang ◽

Wanjing Li ◽

...

Keyword(s):

Red Blood Cells ◽

Whole Blood ◽

Blood Cells ◽

Morphological Changes ◽

Osmotic Fragility ◽

Spherical Shape ◽

Hypoxic Environment ◽

Wide Range ◽

Group A ◽

Cell Membrane Protein

Background: The hypoxic environment stimulates the human body to increase the levels of hemoglobin (HGB) and hematocrit and the number of red blood cells. Such enhancements have individual differences, leading to a wide range of HGB in Tibetans’ whole blood (WB). Study Design: WB of male Tibetans was divided into 3 groups according to different HGB (i.e., A: >120 but ≤185 g/L, B: >185 but ≤210 g/L, and C: >210 g/L). Suspended red blood cells (SRBC) processed by collected WB and stored in standard conditions were examined aseptically on days 1, 14, 21, and 35 after storage. The routine biochemical indexes, deformability, cell morphology, and membrane proteins were tested. Results: Mean corpuscular volume, adenosine triphosphate, pH, and deformability were not different in group A vs. those in storage (p > 0.05). The increased rate of irreversible morphology of red blood cells was different among the 3 groups, but there was no difference in the percentage of red blood cells with an irreversible morphology after 35 days of storage. Group C performed better in terms of osmotic fragility and showed a lower rigid index than group A. Furthermore, SDS-PAGE revealed similar cross-linking degrees of cell membrane protein but the band 3 protein of group C seemed to experience weaker clustering than that of group A as detected by Western Blot analysis after 35 days of storage. Conclusions: There was no difference in deformability or morphological changes in the 3 groups over the 35 days of storage. High HGB levels of plateau SRBC did not accelerate the RBC change from a biconcave disc into a spherical shape and it did not cause a reduction in deformability during 35 days of preservation in bank conditions.

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Glycophorin A-based exclusion of red blood cells for flow cytometric analysis of platelet glycoprotein expression in citrated whole blood

Clinical Chemistry and Laboratory Medicine (CCLM) ◽

10.1515/cclm-2020-0014 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Christina Berens ◽

Johannes Oldenburg ◽

Bernd Pötzsch ◽

Jens Müller

Keyword(s):

Red Blood Cells ◽

Expression Analysis ◽

Whole Blood ◽

Blood Cells ◽

Control Sample ◽

Small Sample ◽

Platelet Glycoprotein ◽

Glycophorin A ◽

Giant Platelets ◽

Platelet Counts

AbstractObjectivesAnalysis of platelet glycoprotein (GP) expression by flow cytometry is applied for diagnostic confirmation of GP-associated thrombocytopathies. While platelet-rich plasma may be used for distinct identification of target events, this strategy is not feasible for small sample volumes or for patients showing low platelet counts and/or giant platelets. However, also the use of whole blood (WB) is hampered by the difficulty to discriminate platelets from red blood cells (RBC) in such patients. To circumvent these limitations, we evaluated the feasibility of a RBC gating-out strategy.MethodsIn addition to platelet GPIb, GPIIa/IIIa, as well as P-selectin (CD62P), citrated whole blood (CWB) samples were stained for RBC-specific glycophorin A (CD235a). CD235a-negative platelet events were further discriminated by forward-/side-scatter characteristics and platelet GP expressions analyzed relative to that of a healthy control sample processed in parallel.ResultsEstablished reference intervals allowed for clear identification of decreased GPIIb/IIIa- or GPIb expression pattern in samples of patients with confirmed Glanzmann thrombasthenia or Bernard–Soulier syndrome, respectively. It could be shown that the analysis of 2,500 platelet events is sufficient for reliable GP expression analysis, rendering the proposed method applicable to samples with low platelet counts.ConclusionsThis study demonstrates the feasibility of CD235a-based exclusion of RBC for platelet GP expression analysis in CWB. In contrast to direct staining of platelet-specific antigens for target identification, this indirect gating out approach is generally applicable independent of any underlying platelet GP expression deficiency.

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