Evaluation of red blood cells stored at -80 degrees C in excess of 10 years.

Author(s): Lecak J, Scott K, Young C, Hannon J, Acker JP

Publication: Transfusion, 2004, Vol. 44, Page 1306-13

PubMed ID: 15318853 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of pre-freeze storage duration at 4 degrees C, duration of storage at -80 degrees C in 40% glycerol (5.1-8.4 years versus 10.5-22.9 years) and 0 h or 24 h post-thaw storage at 4 degrees C on markers of red blood cell (RBC) quality. RBCs from healthy blood donors were collected in citrate phosphate dextrose-adenine 1 (CPDA-1) from 1980-1992 (this included 9 of the 10 specimens stored >10 years) and were collected in citrate phosphate double dextrose (CP2D) and then stored in additive solution 3 (AS3) from 1992-1998 (all of the specimens stored <10 years and 1 specimen that was stored 10.5 years). RBCs in CPDA-1 were stored at 4 degrees C for 1-7 days before freezing and specimens in CP2D/AS3 were stored for 3-18 days at 4 degrees C before freezing. For freezing, a glycerol-lactate solution was added to achieve a final concentration of 40% glycerol, and specimens were frozen in metal containers at -80 degrees C. Specimens were thawed at 36-38 degrees C and washed prior to assay. RBCs were assayed immediately after thaw and again after 24 h at 4 degrees C.

   Summary of Findings:

   Post-wash recovery, supernatant Hb, MetHb, MCV, MCH, and ATP levels were comparable in RBCs stored for <10 years or >10 years, and the authors report that frozen storage duration had no effect on RBC morphology. However, levels of 2,3 DPG were lower in specimens stored for <10 years at -80 degrees C than those stored for >10 years both immediately after thaw (p<0.005) and after 24 h post-thaw storage at 4 degrees C (p<0.005). MCHC was lower in specimens stored for >10 years at -80 degrees C than those stored for <10 years after 24 h post-thaw storage at 4 degrees C (p=0.047), but not immediately post-thaw. Immediately post-thaw, MCF was lower in specimens stored <10 years at -80 degrees C than those stored >10 years (p<0.047). MCV, MCH, and MCHC were comparable immediately post-thaw and after 24 h post-thaw storage at 4 degrees C, but ATP levels declined significantly after 24 h post-thaw storage at 4 degrees C (frozen <10 years, P=0.024; frozen >10 years, P=0.018). Pre-freeze refrigerated storage duration was not predictive of post-wash recovery of RBCs or ATP levels, but 2,3 DPG levels declined with increasing pre-freeze storage duration, and a non-significant interaction between pre-freeze refrigerated storage duration and frozen storage duration was observed for post-thaw RBC recovery and ATP levels.

   Biospecimens

   • Cell - Red Blood Cells

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal

   Platform:

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

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0 h post-thaw 24 h post-thaw 1-18 days pre-freeze 5.1-8.4 years frozen 10.5-22.9 years frozen
   Storage	Storage temperature	4 degrees C -80 degrees C
   Storage	Short-term storage solution	CP2D/AS3 CPDA-1

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