Buffy coat (top/bottom)‐ and whole‐blood filtration (top/top)‐produced red cell concentrates differ in size of extracellular vesicles

Bicalho, Beatriz; Pereira, André; Acker, Jason P.

doi:10.1111/vox.12272

Cited by 37 publications

(51 citation statements)

References 17 publications

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“…It has been indicated that RBC membrane changes occur ex vivo and that there is a reduction in ATP and an increase in percent hemolysis, which are clearly associated with RBC membrane changes and microvesiculation [31,51,56,57,58]. This supports our findings that a positive correlation between EVs and hemolysis and a negative correlation between EVs and the level of ATP or deformability parameters (El max ) were observed.…”

Section: Discussionsupporting

confidence: 92%

“…It has been suggested that the characteristics of EVs can vary among the blood products due to the variation in the blood processing methods [51,52]. An article by Bakkour et al [19] highlighted some feasible reasons to explain the variation in EV characteristics between RCF and WBF RCCs, including variability in the temperature and the lengths of pre-processing storage time as well as leukoreduction technique.…”

Section: Discussionmentioning

confidence: 99%

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Characteristics of Extracellular Vesicles in Red Blood Concentrates Change with Storage Time and Blood Manufacturing Method

Almizraq

Holovati

Acker

2018

Transfus Med Hemother

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Background: Extracellular vesicles (EVs) in blood products are potential effectors of inflammation and coagulation after transfusion. The aim of this study was to assess the impact of different blood manufacturing methods and duration of hypothermic storage on the EV subpopulations in relation to other in vitro quality parameters of red blood cell concentrate (RCC) products. Methods: RCCs were produced using whole blood filtration (WBF) or red cell filtration (RCF) (n = 12/method), refrigerated for 43 days, and evaluated for EV size profile and concentration, red cell deformability, ATP and 2,3-DPG, hemolysis, and hematological indices. Results: The total number of EVs increased significantly with storage in both methods, and WBF-RCCs contained the higher numbers of EVs compared to RCF-RCCs. The concentration of small EVs was greater in WBF-RCCs versus RCF-RCCs, with difference between the two methods observed on day 43 of storage (p = 0.001). Throughout storage, significant decreases were identified in ATP, 2,3-DPG, and EI_max, while an increase in hemolysis was observed in both RCC products. Conclusion: The dynamic shift in the size and concentration of the EV subpopulations is dependent on the blood manufacturing method and length of storage. Better understanding of the potential clinical implications of these heterogeneous populations of EVs are needed.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Characteristics of Extracellular Vesicles in Red Blood Concentrates Change with Storage Time and Blood Manufacturing Method

Almizraq

Holovati

Acker

2018

Transfus Med Hemother

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“…3), suggesting that in the RCCs there was a mixture of exosomes (50-100 nm) and plasma membrane microvesicles (ectosomes, 200-250 nm) changing in proportion during storage [20], which is consistent with the accumulation of RBC EVs (ectosomes) during storage (fig. 1).…”

Section: Resultsmentioning

confidence: 71%

“…Flow cytometry and the use of Trucount tubes (BD Biosciences, San Jose, CA, USA) for glycophorin A-positive (CD235a+) extracellular vesicle (EV) quantification was performed as elsewhere described [19,20]. Briefly, RBCs were sampled from the units, and 5 µl were diluted with HEPES Ca 2+ (2.5 mmol/l) buffered saline, pH 7.4, mixed with 5 µl each anti-CD235a fluorescein isothiocyanate (FITC), anti-CD47 phosphatidylethanolamine (PE) and Annexin V-APC, and transferred to Trucount tubes (BD Biosciences).…”

Section: Methodsmentioning

confidence: 99%

“…Tubes were incubated for 15 min in the dark at room temperature and then vortexed before analysis using a FACSCalibur flow cytometer (BD Biosciences). Forward scatter and side scatter channels were set to log scale to accommodate the visualization of cells and vesicles in a single panel as shown elsewhere [20]. Acquisition was set up on the basis of time (3 min) without gating.…”

Section: Methodsmentioning

confidence: 99%

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Blood Bag Plasticizers Influence Red Blood Cell Vesiculation Rate without Altering the Lipid Composition of the Vesicles

Bicalho

Serrano

Pereira

et al. 2015

Transfus Med Hemother

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Background: Polyvinyl chloride (PVC) plasticized with di(2-ethylhexyl) phthalate (DEHP) is commonly used for blood collection and storage. DEHP has protective effects on RBC membranes, but is also a toxin. Methods: A paired study was conducted to investigate the influence of DEHP and two alternative plasticizers, 1,2-cyclohexane-dicarboxylic acid diisononyl ester (DINCH) and n-butyryl-tri-n-hexyl citrate (BTHC), on the preservation of RBCs stored for 42 days in PVC pediatric bags. The RBC membrane was evaluated for supernatant hemoglobin (Hb), release of extracellular microvesicles (EVs), osmotic fragility, deformability, and lipid composition. Results: In BTHC-plasticized bags, the supernatant Hb increase during storage was 2 times greater than in DINCH- and DEHP-plasticized bags. By day 21, EV concentrations had doubled from day-5 levels in DINCH- and DEHP-, and trebled in BTHC-plasticized bags. RBC mean cell volumes were greater in BTHC- than in DINCH- or DEHP-plasticized bags (p < 0.001). Osmotic fragility differed significantly among plasticizers (p < 0.01). After day 21, RBC deformability decreased in all, but to a greater extent in the bags with BTHC. Phospholipid composition of RBCs and EVs was not different among plasticizers. Conclusion: Membrane stabilization capacity differed among the plasticizers. RBC in BTHC bags stored more poorly, while DEHP and DINCH bags offered better protection against vesiculation, osmotic stress, and Hb loss.

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The role of extracellular vesicles from stored RBC units in B lymphocyte survival and plasma cell differentiation

Gao

Jin

Tan

et al. 2020

Journal of Leukocyte Biology

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Extracellular vesicles (EVs) are small, double-membrane vesicles derived from erythrocytes, leukocytes, platelets, and cells of multiple tissues under physiologic or pathologic conditions. The role of EVs in stored RBC units is of great interest with respect to transfusion-related immunomodulation. The current study focuses on the quantity of EVs isolated from stored RBC units and their action on B cell-mediated immune responses. The in vitro experiment demonstrated that EVs exhibited a negative role in B cell survival, plasmacytic differentiation, and class switch recombination under LPS stimulation. Furthermore, LPS-induced antibody production was significantly decreased after EVs injection in vivo. Biochemical analysis revealed that EVs hampered the expression of Blimp-1 and IRF4 and the activation of NF-B pathway in LPS-primed B cells. Overall, these data imply a vital role for EVs isolated from RBC units in B cell-mediated immune responses.

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Buffy coat (top/bottom)‐ and whole‐blood filtration (top/top)‐produced red cell concentrates differ in size of extracellular vesicles

Cited by 37 publications

References 17 publications

Characteristics of Extracellular Vesicles in Red Blood Concentrates Change with Storage Time and Blood Manufacturing Method

Characteristics of Extracellular Vesicles in Red Blood Concentrates Change with Storage Time and Blood Manufacturing Method

Blood Bag Plasticizers Influence Red Blood Cell Vesiculation Rate without Altering the Lipid Composition of the Vesicles

The role of extracellular vesicles from stored RBC units in B lymphocyte survival and plasma cell differentiation

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