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Exposure of cryopreserved red cell concentrates to real-world transient warming events has a negligible impact on quality.

Author(s): Kurach J, Brandon-Coatham M, Olafson C, Turner TR, Phan C, Yazdanbakhsh M, Osmani R, Ehsani-Moghaddam B, Clarke G, Acker JP

Publication: Transfusion, 2024, Vol. 64, Page 2353-2363

PubMed ID: 39485279 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study compared the quality of red cell concentrates (RCC) that experienced transient warming events (≥-65°C) due to freezer failures of 34 min and/or 48 h during long-term storage and in control RCCs with no known warming events during storage. Specimens were compared immediately after thaw and after 1, 7, and 14 days of hypothermic storage. Forty cryopreserved RCC that were rejected from Canadian Blood Services locations across Canada due to age or warming were obtained. Specimens had been stored for 3.4-17.2 years. Specimens were classified based on the number of times that they had experienced temperatures > -65°C due to freezer failures during storage: ten specimens were stored for the same duration but never warmed above > -65°C (storage duration 3.4-11.6 y), five specimens spent 34 min at > -65°C (max temp -49°C, storage duration 5.2-11.7 y), twenty-three specimens spent 48 h at > -65°C (max temp -30°C, storage duration 7.5-17.4 y), and two specimens spent both the 34 min and 48 h at > -65°C (storage duration 11.8-11.9 y). Thirty-eight of the specimens had been manually glycerolized, but two of the specimens that spent 34 min at > -65°C were glycerolized using an ACP 215. Specimens were thawed and deglycerized using an ACP 215, resuspended and sampled aseptically on day 0, 1, 7 and 14 of hypothermic storage. Hemoglobin and mean corpuscular volume were measured using a DxH 520 hematology analyzer. Extracellular potassium levels were measured after centrifugation at 2200 g for 10 min at 4°C using a Roche Cobas c503. Spun hematocrit and RBC hemolysis were analyzed by autoanalyzer. Oxygen affinity was assayed using a Hemox-Analyzer Model B. ATP concentration was measured determined using the DiaSys Diagnostic Systems enzyme-assay. Red blood cell morphology was scored using fixed and stained blood smears. Osmotic fragility was assessed by exposing red blood cells to 1.0–9.0 g/L saline and determining the concentration that produces 50% hemolysis (mean corpuscular Fragility, MCF).  Red blood cell deformability was assessed with and without exposure to an osmotic gradient. 

   Summary of Findings:

   Hemoglobin level, hematocrit and red blood cell hemolysis met the Canadian Standard Association standards in 100% of the RCC that had experienced transient warming events (34 min,   48 h, or 34 min and 48 h at > -65°C), but  90%, 100% and 100% of control specimens that had not experienced transient warming met the Canadian Standard Association standards for hemoglobin level, hematocrit and red blood cell hemolysis, respectively. Further, red blood cell hemolysis, hematocrit, mean corpuscular volume were not affected by transient warming events at any of the timepoints evaluated (0-14 days post-thaw). In contrast, the specimens that experienced two transient warming events had lower osmotic hemolysis values at each timepoint post-thaw than specimens that did not experience any warming events. ATP and p50 levels declined with post-thaw storage, and on day 7 were lower in specimens that experienced a 48 h transient warming event than those that only experienced the 34 min event (P<0.001 and P<0.05, respectively). Extracellular potassium levels increased and morphological index scores decreased with storage, regardless of warming events.  While overall deformability of red blood cells was not significantly affected by warming events, ΔElongation was lower on day 14 of storage and EImax was lower at all timepoints examined in specimens that experienced both warming events than those that experienced just one. EI_hyper was lower in specimens that experienced two warming events than in control specimens or specimens that only experienced 48 h of warming; but on day 1 post-thaw, EI_hyper was lower in specimens that experienced the 48 h transient warming event and higher in control specimens than in specimens that experience the 34 min warming event (P<0.001 and P<0.05, respectively). A similar trend was observed for the Osmoscan hyper index.

    

   Biospecimens

   • Cell - Red Blood Cells

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Hematology/ auto analyzer
   Small molecule	Enzyme assay
   Morphology	Light microscopy
   Electrolyte/Metal	Clinical chemistry/auto analyzer
   Protein	Hematology/ auto analyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage conditions	Single transient warming event of 34 min Single transient warming event of 48 h Transient warming event of 34 min and 48 h No known transient warming events
   Storage	Storage duration	Day 0 1 day 7 days 14 days

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