Altered processing of thawed red cells to improve the in vitro quality during postthaw storage at 4°C

Hoehn

Jernigan

et al. 2016

Shock

Background During storage, packed red blood cells (pRBCs) undergo a number of biochemical, metabolic and morphologic changes, collectively known as the “storage lesion”. We aimed to determine the effect of cryopreservation on the red blood cell storage lesion compared to traditional 4°C storage. Methods Previously cryopreserved human packed red blood cells were compared to age matched never frozen packed red blood cells obtained from the local blood bank. The development of the red cell storage lesion was evaluated after 7, 14, 21, 28, and 42 days of storage at 4°C in AS-3 storage medium. We measured physiological parameters including cell counts, lactic acid and potassium concentrations as well as signs of eryptosis including loss of phosphatidylserine (PS) asymmetry, microparticle production and osmotic fragility in hypotonic saline. Results Compared to controls, previously cryopreserved pRBC at 7 days of storage in AS-3 showed lower red cell counts (3.7 vs 5.3 ×10^6 cells/uL, p(<0.01), hemoglobin (12.0 vs 16.5 g/dL, p<0.01), hematocrit (33.0 vs 46.5%, p<0.01), and pH (6.27 vs 6.72, p<0.01). Over 28 days of storage, storage cryopreserved pRBC developed increased cell free hemoglobin (0.7 vs 0.3 g/dL, p<0.01), greater PS exposure (10.1 vs 3.3%, p<0.01), and microparticle production (30,836 vs 1,802 MP/uL, p<0.01). Previously cryopreserved cells were also less resistant to osmotic stress. Conclusion The red blood cell storage lesion is accelerated in previously cryopreserved pRBC after thawing. Biochemical deterioration of thawed and deglycerolized red cells suggests that storage time prior to transfusion should be limited in order to achieve similar risk profiles as never frozen standard liquid storage pRBC units.

Section: Discussionmentioning

confidence: 99%

Previous Cryopreservation Alters the Natural History of the Red Blood Cell Storage Lesion

Hoehn

Jernigan

et al. 2016

Shock

“…ATP and 2,3-DPG concentrations of the erythrocytes were measured using a previously published method with slight modifi cation (Lagerberg et al 2007). Each thawed erythrocyte sample (0.2 ml) was added to 1 ml of perchloric acid ( …”

Section: Erythrocyte Metabolite Assaymentioning

confidence: 99%

“…Th e optical densities (OD) of total hemoglobin (tHb) and supernatant hemoglobin (sHb) were read spectrophotometrically at a wavelength of 540 nm (Model 2020; Anthos Labtec Instruments, Wals, Salzburg, Austria). Percentages of hemolysis and cell survival rate were calculated using the following formulas (Lagerberg et al 2007):…”

Section: Erythrocyte Recovery Testmentioning

confidence: 99%

Influence of a static magnetic field on the slow freezing of human erythrocytes

Lin

International Journal of Radiation Biology

Lee

et al. 2012

“…3–6 Other studies have identified a host of molecular and biochemical changes that occur during packed red blood cell (pRBC) storage, collectively known as the red blood cell storage lesion, as the etiology of massive transfusion related morbidity. 6–8 Although these changes are documented in the literature, there is little understanding as to how agents in donor blood interacts with the transfusion recipient to cause harm.…”

Section: Introductionmentioning

confidence: 99%

Erythrocyte-Derived Microparticles Activate Pulmonary Endothelial Cells in a Murine Model of Transfusion

Kim

Seitz

et al. 2017

Shock

Erythrocyte-derived microparticles (MPs) are sub-micrometer, biologically active vesicles shed by red blood cells as part of the biochemical changes that occur during storage. We hypothesized that MPs from stored red blood cells would activate endothelial cells. MPs from aged murine packed red blood cells (pRBCs) were isolated and used to treat confluent layers of cultured endothelial cells. Endothelial expression of leukocyte adhesion molecules, ELAM-1 and ICAM-1, and inflammatory mediator, IL-6, were evaluated at 0.5, 6, 12, and 24 hours of treatment. Healthy C57BL/6 mice were transfused with a MP suspension and lung sections were analyzed for adhesion molecules and sequestered interstitial leukocytes. Increased levels of ELAM-1 and ICAM-1 were found on cultured endothelial cells 6 hours after MP stimulation (6.91 vs 4.07 relative fluorescent intensity, RFI, p<0.01, and 5.85 vs 3.55 RFI, p=0.01, respectively). IL-6 in cell culture supernatants was increased after 12 hours of MP stimulation compared to controls (1.24 vs 0.73 ng/ml, p=0.03). In vivo experiments demonstrated that MP injection increased ELAM-1 and ICAM-1 expression at 1 hour (18.56 vs 7.08 RFI, p<0.01, and 23.66 vs 6.87 RFI, p<0.01, respectively) and caused increased density of pulmonary interstitial leukocytes by 4 hours of treatment (69.25 vs 29.25 cells/HPF, p<0.01). This series of experiments supports our hypothesis that erythrocyte-derived MPs are able to activate pulmonary endothelium, leading to the pulmonary sequestration of leukocytes following the transfusion of stored pRBCs.