The effects of long-term storage of human red blood cells on the glutathione synthesis rate and steady-state concentration.

Author(s): Whillier S, Raftos JE, Sparrow RL, Kuchel PW

Publication: Transfusion, 2011, Vol. 51, Page 1450-9

PubMed ID: 21251007 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 refrigerated storage duration and storage additives on TFG synthesis rate and concentration, pH, ATP levels, thiol concentration and HbA1c in RBC specimens from healthy volunteers. Blood was processed using the buffy-coat method, and RBCs were suspended in saline-adenine-glucose-mannitol (SAGM) and leukoreduced by filtration. 65 mL of RBCs were placed into 4 pediatric bags with 2.5% SAGM (Control), N-acetylcysteine with alanine, a-ketoglutarate, and glycine in SAGM (Amino acid), aminoguanidine in SAGM, or glyoxal in SAGM. The effects of rejuvenation were assayed by treatment with rejuvenation solution. ATP, TFG and GSH synthesis rate were assayed during storage (20 min-3h) at 37 degrees C.

   Summary of Findings:

   TFG level remained constant until week 3 of refrigerated storage and then declined (p<0.0001), with the exception of amino acid treated RBCs, where TFG levels remained constant. TFG was lower in rejuvenated RBCs refrigerated for 1 or 3 weeks than in unrejuvenated RBCs (p=0.04 and p=0.05, respectively). Only non-significant declines in thiol concentrations were noted in control aminoguanidine and glyoxal treated RBCs during refrigerated storage. With the exception of glyoxal treated RBCs which had lower TFG synthesis after 3 weeks of refrigerated storage, TFG synthesis was constant during the first 3 weeks of refrigeration of all other RBC specimens, but then declined by week 6(p<0.0001), except in the presence of amino acids or rejuvenation solution. Glyoxal treated RBCs showed lower TFG synthesis than control RBCs after 3 and 6 weeks of refrigeration (p<0.05, both). The pH of RBCs decreased over the first 3 weeks of refrigeration (p=0.004) and was lower still after 6 weeks (p<0.0001), regardless of treatment. With the exception of RBCs in rejuvenation solution, the ATP content declined with refrigeration of RBCs (p<0.05). HbA1c levels were unaffected by refrigerated storage of RBCs, regardless of treatment. Modeling identified the cause of the decline in GSH to be lower substrate and ATP concentrations.

   Biospecimens

   • Cell - Red Blood Cells

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	Hematology/ auto analyzer
   Small molecule	Bioassay
   Small molecule	Spectrophotometry
   Peptide	Colorimetric assay

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Short-term storage solution	SAGM SAGM and GSH precursor amino acids SAGM and aminoguanidine SAGM and glyoxal Rejuvenation solution
   Storage	Storage duration	1 week 3 weeks 6 weeks

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