A novel model of red blood cell storage and posttransfusion in vivo survival

Center for Transfusion and Cellular Therapies, Department of Pathology and Laboratory Medicine, Emory University School of Medicine,Woodruff Memorial Building Suite 7107A, 101Woodruff Circle, Atlanta, GA 30322, USA.
Transfusion (Impact Factor: 3.23). 08/2009; 49(8):1546-53. DOI: 10.1111/j.1537-2995.2009.02173.x
Source: PubMed

ABSTRACT Storage of red blood cells (RBCs) is necessary for an adequate blood supply. However, reports have identified potential negative sequelae of transfusing stored RBCs. An animal model would be useful to investigate the pathophysiology of transfusing stored RBCs. However, it has been reported that storage of rat RBCs in CPDA-1 resulted in an unexpected sudden decline in posttransfusion survival. A mouse model of RBC storage and transfusion was developed to assess survival kinetics of mouse RBCs.
RBCs expressing green fluorescent protein were collected in CPDA-1, filter leukoreduced, adjusted to a 75% hematocrit, and stored at 4°C. At weekly intervals, stored RBCs were transfused into C57BL/6 recipients. RBC survival was measured by flow cytometry and chromium-51 labeling. Phosphatidylserine externalization and CD47 expression was also evaluated.
Mean 24-hour survivals of transfused RBCs were 99, 91, 64, 54, 30, and 18% after 0, 7, 14, 21, 28, and 35 days of storage, respectively. Stored RBCs showed an initial rapid clearance with subsequent extended survival. Increased surface phosphatidylserine and decreased CD47 expression were also observed.
Mouse RBCs showed a progressive decline in survival, as a function of storage time, unlike the precipitous loss of viability reported for rat RBCs. Moreover, changes in the measured surface markers were analogous to trends reported for human RBCs. Together, these findings provide an initial characterization of a novel mouse model of RBC storage with the potential to serve as an experimental platform for studying the pathophysiologic consequences of transfusing stored RBCs.

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Available from: Steven L Spitalnik, Sep 29, 2015
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    • "The effects resulting from storage lesions that may be associated in a reduction in the quality of blood products is a concern for all blood banks. Whether storage lesions result in adverse clinical effects has been controversially discussed in the literature (Ho et al., 2003; Raghavan & Marik, 2005; van de Watering & Brand, 2008; Gilson et al., 2009; Vandromme et al., 2009; Zimrin & Hess, 2009; van de Watering, 2011b; Wang et al., 2012; Aubron et al., 2013; van de Watering, 2013 "
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    ABSTRACT: The objective of this study was to elucidate whether complement activation occurs during the storage of RBCs in newly formulated PAGGS-M storage medium. The reason for red blood cell (RBC) storage lesions is not yet fully understood. The contribution of complement to RBC storage lesion has not been extensively characterised. We investigated the surface expression of CD35, CD55, CD59 and CD47, as well as deposition of C3d, using flow cytometry over a storage period of up to 42 days on a weekly basis. C3d and the immunoglobulins IgG, IgM and IgA were additionally investigated via the direct antiglobulin test (DAT). The effect of contact with homologous serum for 30 min at 37 °C was also performed for C3d and CD35 and is subsequently termed as a 'transfusion simulation (TS)'. A weak but significant increase of C3d was observed prior to TS (anova P = 0·0103), whereas a stronger increase from 74·0 ± 12·4 to 101·2 ± 9·7 was observed post-TS (anova; P < 0·0001). These findings were confirmed by the DAT. CD35, CD55 and CD47 demonstrated a decrease in their expression over storage time (anova; P < 0·0001 each). The majority of changes occurred following 14 days. There was neither a decrease of CD59 observed nor an increase of IgG, IgM and IgA. RBCs are becoming increasingly susceptible to spontaneous complement deposition following TS, which might be associated with the decrease of C35 and CD55 by proteolytic cleavage and vesiculation during storage. As the impact of storage lesions is rather controversial, institutions involved in blood collection and administration of blood products should focus on carrying out research on the prevention of storage lesions. © 2014 British Blood Transfusion Society.
    Transfusion Medicine 12/2014; 24(6). DOI:10.1111/tme.12166 · 1.65 Impact Factor
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    • "Recently, inventive efforts to study the post-transfusion survival of RBC in model animal systems have revealed important information related to the storage lesion and transfusion-related reactions [26]–[28]. Using such an established mouse model of blood banking [27], [28] we evaluated the impact of 5-HT supplementation on the 24h post-transfusion survival of stored RBC. In order to closely mimic the aging process undergone by human RBC during hypothermic storage, mouse RBC were collected in the conventional human storage solution CPDA-1, stored at 4°C after removal of the leukocytes by filtration, stained with CFDA-SE and transfused to a recipient to follow their post-transfusion survival. "
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    ABSTRACT: Serotonin (5-HT) is a monoamine originally purified from blood as a vasoactive agent. In nonneuronal tissues, its presence is linked with the expression of tryptophan hydroxylase 1 (TPH1) that catalyzes the rate-limiting step of its synthesis. Targeted disruption in mice of the TPH1 gene results in very low levels of circulating 5-HT. Previous analysis of the TPH1 knockout (TPH1(-/-)) mouse revealed that they develop a phenotype of macrocytic anemia with a reduced half-life of their circulating red blood cells (RBC). In this study, to establish whether the observed reduced half-life of TPH1(-/-) RBC is an intrinsic or an extrinsic characteristic, we compared their survival to RBC isolated from wild-type mice. Both in vivo and in vitro data converge to demonstrate an extrinsic protective effect of 5-HT since presence of 5-HT in the RBC environment protects RBC from senescence. The protective effect played by 5-HT is not mediated through activation of a classical pharmacological pathway as no 5-HT receptors were detected on isolated RBC. Rather, 5-HT acts as an effective antioxidant since reduction of 5-HT circulating levels are associated with a decrease in the plasma antioxidant capacity. We further demonstrate a link between oxidation and the removal of damaged RBC following transfusion, as supplementation with 5-HT improves RBC post-transfusion survival in a mouse model of blood banking.
    PLoS ONE 12/2013; 8(12):e83010. DOI:10.1371/journal.pone.0083010 · 3.23 Impact Factor
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    • "A progressive decrease in the CD47 expression level has been observed on human erythrocytes stored under blood bank conditions at +4°C for more than 14 days [107–109] and in a mouse model of erythrocyte storage which tried to mimic human blood bank conditions [110]. However, the magnitude of reduction in CD47 expression was rather different, from a modest reduction of up to 6% at day 42 of storage [107], around 30% reduction on day 28 of storage [108], to a more than 50% reduction on day 14 of storage [109]. "
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    01/2013; 2013(5):614619. DOI:10.1155/2013/614619
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