Metabolomic analysis of platelets during storage: a comparison between apheresis- and buffy coat-derived platelet concentrates.
Author(s): Paglia G, Sigurjónsson ÓE, Rolfsson Ó, Hansen MB, Brynjólfsson S, Gudmundsson S, Palsson BO
Publication: Transfusion, 2015, Vol. 55, Page 301-313
PubMed ID: 25156572 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine if the method of platelet (plt) preparation alters plt activation and metabolic activity during storage.
Conclusion of Paper
A metabolic shift occurred between day 3 and 4 of storage in both buffy-coat-derived and apheresis plts. Prior to this shift, apheresis-derived plts had higher levels of activation markers and more glycolysis and tricarboxylic acid (TCA) cycle activity, but after the shift, rates were comparable. The activation of apheresis-derived plts was variable, resulting in more variability in the metabolic profile.
Studies
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Study Purpose
The purpose of this study was to determine if room temperature storage effects the metabolic profile of buffy-coat-derived and apheresis plts differently. Plts were obtained using the buffy coat method from the blood of 8 healthy donors and were obtained by apheresis from 6 other healthy donors. Plts were stored in T-Sol in a shaking incubator at 22°C. More than 150 variables were measured at each timepoint.
Summary of Findings:
During storage of plts, potassium, lactate, and partial pressure oxygen (PO2) increased, and partial pressure carbon dioxide (PCO2), glucose, and ATP decreased, regardless of plt collection method. However, compared to buffy-coat-derived plts, the rate of glucose consumption was faster in apheresis plts during the first 3 days of storage, and apheresis plts had lower pH from day 1 to day 7 and less lactate production between day 7 and day 10. Markers of plt activation, including the percentage of cells positive for CD41 alone or with Annexin V, CD62P or CD63, soluble CD40 ligand, P-selectin, and the percentage of cells with depolarized mitochondrial membranes, increased during storage of both apheresis and buffy-coat-derived plts, but the increase in activation was faster in apheresis plts than buffy-coat-derived plts. The activation marker, soluble P-selectin, was strongly correlated with external levels of malate, glucose, lactate and hypoxanthine in buffy-coat-derived (r=0.81, r=0.86, r=0.90, and r=0.94, respectively) and apheresis (r=0.85, r=0.86, r=0.87 and r=0.90, respectively) plts. Principle component analysis (PCA) and heatmap analysis identified a shift in the metabolic profile between day 3 and 4 of storage, for both buffy-coat-derived and apheresis plts, PCA identified a second shift on day 7 only in apheresis plts. More variability was also noted among apheresis specimens than buffy-coat specimens and was attributed to the more rapid plt activation and variability in activation. The main pathways responsible for the metabolic shift between day 3 and day 4 were glutathione metabolism, purine metabolism, glycolysis, TCA cycle, pentose phosphate pathway, lipid metabolism, and nicotinamide metabolism. Importantly, during the first phase (day 0-3), apheresis plts had faster consumption and excretion rates (particularly glycolysis and TCA cycle) than buffy-coat-derived plts, but during the second phase, the metabolic profiles of buffy-coat-derived and apheresis plts were very similar. Flow cytometry confirmed that apheresis-derived plts were more likely to be active during the early storage period than buffy-coat-derived plts, and that buffy-coat-derived plt activation was more gradual.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Small molecule Clinical chemistry/auto analyzer Small molecule ELISA Small molecule Spectrophotometry Small molecule LC-MS or LC-MS/MS Gas Clinical chemistry/auto analyzer Cell count/volume Hematology/ auto analyzer Protein ELISA Carbohydrate Clinical chemistry/auto analyzer Electrolyte/Metal Clinical chemistry/auto analyzer Cell count/volume Flow cytometry Protein Clinical chemistry/auto analyzer Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Method of fluid acquisition Apheresis
Venipuncture
Storage Time at room temperature 0 days
1 day
2 days
3 days
4 days
5 days
6 days
7 days
10 days
Biospecimen Aliquots and Components Blood processing method Buffy coat method
Apheresis