scholarly journals Folate Forms in Red Blood Cell Lysates and in Intact Washed Red Blood Cells Are Stable for a Few Days During Refrigerated Storage

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1799-1799
Author(s):  
Zia Fazili ◽  
Neelima Papadugula ◽  
Phuong Ngac ◽  
Christine M Pfeiffer
Keyword(s):  
Ascorbic Acid ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Solid Phase ◽  
Storage Conditions ◽  
Sample Type ◽  
Refrigerated Storage ◽  
Funding Sources ◽  
Edta Blood ◽  
Time Point

Abstract Objectives We investigated whether folate forms are stable in washed red blood cells (RBC) stored refrigerated for up to 9 days relative to RBC-lysates in ascorbic acid under the same storage conditions. Methods We prepared washed RBCs from freshly collected EDTA blood (n = 6 donors). For RBC-lysate 1, we diluted RBCs with saline (1/2 dilution), mixed well and prepared lysates (1/11 dilution) with 1% ascorbic acid (2 vials/time point; n = 12 vials/donor) for storage at 4°C for ≤ 9 days (baseline, 1, 2, 3, 6, and 9). For RBC-lysate 2, we aliquoted ∼0.5 mL washed RBCs (1 vial/time point; n = 5 vials/donor) for storage at 4°C for ≤9 days (1, 2, 3, 6, and 9). When refrigerated storage of samples was completed, RBC lysate 1 samples were frozen at −70°C, while the washed RBCs were diluted with saline (1/2 dilution), mixed well and RBC-lysates 2 were prepared (1/11 dilution) with 1% ascorbic acid (2 vials/time point; n = 10 vials/donor). All samples were stored at −70°C until analysis. At the time of analysis, we processed samples (2 replicates/time point; n = 2 days) for folate polyglutamate deconjugation using recombinant exo γ-glutamyl hydrolase, and conducted sample clean-up by automated solid phase extraction prior to analysis by LC-MS/MS. Results We found negligible losses of major folate forms after overnight refrigerated storage, and folate losses gradually increased over time (∼5% by day 6). The loss of 5-methyltetrahydrofolate and total folate (mean ± SD) after 2 days of storage for RBC lysate 1 was 1.2% ± 1.5% and 1.0% ± 1.3%, and for RBC-lysate 2 was 2.2% ± 1.1% and 2.7% ± 1.9%, respectively. The baseline concentration of non-methyl folate (sum of minor folate forms: 5-formyltetrahydrofolate [<LOD], tetrahydrofolate, and 5, 10-methenyltetrahydrofolate) was small (<3.0 nmol/L) in both sample types, and we noticed a slight increase (∼10%) after overnight storage. 5,10-Methenyltetrahydrofolate appeared to gradually convert to tetrahydrofolate upon storage in RBC-lysate 2 samples. We observed no noticeable changes for MeFox in either sample type. Conclusions Overnight refrigerated storage of RBC-lysates or washed RBCs is feasible and avoids notable losses of folate forms. Prolonged refrigerated storage promotes interconversions of minor folate forms. Funding Sources This work was supported by direct appropriations from U.S. Congress.

Donor Variation In Biological Mediators During Storage Of Packed Red Blood Cells

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3655-3655 ◽  
Author(s):  
Melinda M Dean ◽  
Luke D Samson ◽  
Kelly Rooks ◽  
Jesse Fryk ◽  
Shoma Baidya ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Immune Modulation ◽  
Time Course ◽  
Conflicts Of Interest ◽  
Biological Response ◽  
Storage Lesion ◽  
Packed Red Blood Cells ◽  
Donor Variation ◽  
Time Point

Abstract Introduction During routine storage, packed red blood cells (PRBC) undergo numerous biochemical and biophysical changes collectively referred to as the “RBC storage lesion”. A number of factors reported to accumulate during the routine storage of PRBCs are hypothesized to mediate inflammatory cell responses and contribute to poor patient outcomes following transfusion. In addition, donor variability in red blood cell (RBC) characteristics and onset of the storage lesion has been reported. We investigated changes in levels of potential biological response modifies in the supernatant (SN) of PRBC relevant to storage, and, variance between donations. Methods Cytometric bead array was utilised to quantify a panel of 32 potential biological response modifiers (BRMs) in the SN of PRBC during storage. Potential BRMS were analysed in the SN of 8 leukodepleted PRBC units at weekly intervals (D2, D7, D14, D21, D28, D35, D42). The CBA panel was comprised of soluble(s) CD40 Ligand, sCD62E, sCD62L, sCD14, sCD54 (ICAM-1), sCD106 (VCAM-1), CXCL9, VEGF, Fractalkine (CX3CL1), IL-1β, IL-6, IL-8, IL-10, IL-12p70, TNF-α, MIP-1α, MIP-1β, IP-10, RANTES, sCD62P, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-7, IL-9, IL-13, IFN-α, IFN-γ, angiogenin, MCP-1. Storage related changes were analysed using ANOVA (95% CI). Donor variance was indicated by fold difference and range. “High” sub population of donations compared to remaining donations at each time point using Mann-Whitney (95% CI). Results Of the 32 potential BRMs studied, angiogenin, sCD14, sCD106 (VCAM-1), sCD62L, sCD62P, ICAM-1, IL-1α, IP-10, RANTES and IL-9 were consistently detected in all units throughout the time course. There was no evidence of a storage related increase in these biological mediators during storage of the PRBC, although angiogenin levels significantly declined during storage (P<0.001, ANOVA). Of particular interest, the concentrations of these nine biological mediators varied greatly between the individual PRBC units. ICAM-1, VCAM-1 and IL-1α concentrations each varied 10 fold between units (range 1000 – 10 000 pg/mL for each), sCD14 varied 5 fold (range 20 000 - 100 000 pg/mL), sCD62L varied 4.4 fold (range 9000 – 40 000 pg/mL), and sCD62P varied 6.5 fold (range 200 -1300 pg/ml). In addition, it was apparent that a sub population (3/8) of the units assessed consistently had the highest levels of ICAM-1, sCD106 (VCAM-1), sCD14, sCD62L, IL-1α, sCD62P and angiogenin. For sCD62P, in particular, this “high” sub population had significantly different levels of sCD62P at each time point compared to the other five units (P<0.05 at each time point). The remaining BRMs studied were at the limits of detection (<20 pg/mL) for every unit at each time point, and no storage related changes were evident. Conclusions There was minimal change in the BRMs studied relevant to storage duration of the PRBC units. The most notable differences in the levels of biological mediators present in PRBC SN were due to donor-to-donor variation. These data suggest high levels of BRMs and potential immune modulation in transfusion recipients may be the result of donor-associated differences rather than storage-associated differences in blood components. Disclosures: No relevant conflicts of interest to declare.

Addition of ascorbic acid solution to stored murine red blood cells increases posttransfusion recovery and decreases microparticles and alloimmunization

Transfusion ◽  
2013 ◽  
pp. n/a-n/a ◽  
Author(s):  
Sean R. Stowell ◽  
Nicole H. Smith ◽  
James C. Zimring ◽  
Xiaoyun Fu ◽  
Andre F. Palmer ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Acid Solution ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Ascorbic Acid Solution

Transitory interruption of recommended storage conditions does not cause significant changes in in vitro parameters of leucocyte-depleted red blood cells

Vox Sanguinis ◽  
2010 ◽  
Vol 100 (3) ◽  
pp. 272-278 ◽  
Author(s):  
D. R. Weiss ◽  
S. Fortenbacher ◽  
J. Ringwald ◽  
E. F. Strasser ◽  
R. Zimmermann ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Storage Conditions

scholarly journals Performance of an automated solid-phase red cell adherence system compared with that of a manual gel microcolumn assay for the identification of antibodies eluted from red blood cells

2019 ◽  
Vol 27 (1) ◽  
pp. 1-5
Author(s):  
Rachel H. Finck ◽  
Rebecca J. Davis ◽  
Shih-Mao Teng ◽  
Dennis Goldfinger ◽  
Alyssa F. Ziman ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Solid Phase ◽  
Red Cell ◽  
Cell Adherence

scholarly journals Deformability of Stored Red Blood Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Gregory Barshtein ◽  
Ivana Pajic-Lijakovic ◽  
Alexander Gural
Keyword(s):  
Red Blood Cells ◽  
Cold Storage ◽  
Blood Cells ◽  
Storage Conditions ◽  
Cell Deformability ◽  
Red Cell Deformability ◽  
Peripheral Tissues ◽  
Pass Through

Red blood cells (RBCs) deformability refers to the cells’ ability to adapt their shape to the dynamically changing flow conditions so as to minimize their resistance to flow. The high red cell deformability enables it to pass through small blood vessels and significantly determines erythrocyte survival. Under normal physiological states, the RBCs are attuned to allow for adequate blood flow. However, rigid erythrocytes can disrupt the perfusion of peripheral tissues and directly block microvessels. Therefore, RBC deformability has been recognized as a sensitive indicator of RBC functionality. The loss of deformability, which a change in the cell shape can cause, modification of cell membrane or a shift in cytosol composition, can occur due to various pathological conditions or as a part of normal RBC aging (in vitro or in vivo). However, despite extensive research, we still do not fully understand the processes leading to increased cell rigidity under cold storage conditions in a blood bank (in vitro aging), In the present review, we discuss publications that examined the effect of RBCs’ cold storage on their deformability and the biological mechanisms governing this change. We first discuss the change in the deformability of cells during their cold storage. After that, we consider storage-related alterations in RBCs features, which can lead to impaired cell deformation. Finally, we attempt to trace a causal relationship between the observed phenomena and offer recommendations for improving the functionality of stored cells.

Impact of Genetic Predispositions, Diet, and Environment on Storage of Red Blood Cells

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. SCI-38-SCI-38
Author(s):  
Steven L Spitalnik
Keyword(s):  
Red Blood Cells ◽  
Blood Supply ◽  
Blood Cells ◽  
Conflicts Of Interest ◽  
Ex Vivo ◽  
Inbred Strains ◽  
Dietary Factors ◽  
Beta Thalassemia ◽  
Refrigerated Storage ◽  
Transfusion Therapy

It remains controversial whether or not transfusions of human red blood cells (RBCs), which have been refrigerator-stored for extended time intervals, induce or exacerbate clinically-relevant adverse effects in recipients. The described effects include infection, inflammation, renal dysfunction, thrombosis, and even death. It is also controversial whether such effects are seen in particular patient populations predisposed to these types of complications, or whether they are universal. To this end, multiple bench research studies, clinical/translational studies, and prospective randomized controlled trials are currently investigating these issues. What is not controversial is that refrigerated storage produces the "RBC storage lesion" and that this worsens in proportion to increasing storage time, such that, at the FDA-mandated 42-day outdate, on average, only 75% of transfused donor RBCs are still circulating in the recipient at 24-hours post-transfusion. It is also not controversial that RBCs obtained from some healthy volunteer donors do not store well and, at outdate, have a 24-hour post-transfusion recovery (PTR) of significantly less than 75% (i.e., "poor storers"). Analogously, RBCs from some donors store extremely well and may exhibit virtually 100% 24-hour PTR at outdate (i.e., "super storers"). Understanding the underlying mechanisms for these phenomena would allow implementation of various approaches to improve the blood supply, with an ultimate goal of virtually 100% 24-hour PTR and normal long-term circulatory lifespan of transfused RBCs. This goal is especially relevant in the chronic transfusion setting (e.g., patients with sickle cell anemia or beta-thalassemia). Similar observations have been made about hemolysis ex vivo (i.e., "in the bag") during refrigerated storage, which influences the amount of free hemoglobin infused during the transfusion procedure. There is also evidence that these RBC storage characteristics are stable in "poor" and "super" donors, suggesting that genetic, dietary, and/or environmental factors are involved. Indeed, using inbred strains of mice, there is clear evidence that genetic and dietary factors can significantly influence donor RBC storage quality, when measured as 24-hour PTR and/or ex vivo hemolysis. In addition, human genetic polymorphisms, particularly in racial/ethnic groups enriched for malaria-induced traits, result in abnormal expression or abnormal function of various RBC proteins, including hemoglobin and cytosolic enzymes, such as glucose-6-phosphate dehydrogenase. There is increasing evidence that these polymorphisms can influence the ability of human RBCs to tolerate standard conditions of refrigerated storage. Harnessing this information will result in new methods for improving the quality of the blood supply and, consequently, improving patient responses to transfusion therapy. Disclosures No relevant conflicts of interest to declare.

Sign in / Sign up

Export Citation Format

Share Document