PAS-G supports platelet reconstitution after cryopreservation in the absence of plasma.

Author(s): Johnson L, Reid S, Tan S, Vidovic D, Marks DC

Publication: Transfusion, 2013, Vol. 53, Page 2268-77

PubMed ID: 23347144 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 freezing, storage solution and post-thaw room temperature storage on recovery, quality and activation of buffy-coat derived plts. Citrate-phosphate dextrose (CPD) whole blood was stored at room temperature until processing and plt pooling in SSP+. Plts were stored at room temperature overnight, sampled, hyperconcentrated in DMSO solution, and frozen in a -80 degrees C freezer. Plts were thawed in a 37 degrees C water bath and reconstituted in thawed plasma, PAS-G, or SSP+. Reconstituted plts were then stored for up to 24 h at 22 degrees C with agitation.

   Summary of Findings:

   The pH, ATP, PO2, glucose levels and ADP and collagen-induced aggregation decreased, and MPV, lactate levels and percentage of plts positive for CD62P or CD63 increased with room temperature storage of plts in at least one storage solution. pH, plt volume and viability were lowest, and plt recovery, partial pressure carbon dioxide (PCO2), ADP and collagen-induced aggregation and percentage CD63 positive plts were highest for plts in PAS-G at one or more time points. MPV, bicarbonate and lactate were highest, and PO2 and ADP and collagen-induced aggregation were lowest in plts in plasma at one or more time points. PO2 and plt volume were highest, and PCO2, bicarbonate, glucose and ATP levels were lowest in plts in SSP+ at one or more time points. The percentages of plts expressing GP1b alpha and GPIIb were lower for plts in PAS-G than SSP+ (p<0.05). Plt storage solution did not affect HSR or extracellular LDH. Freezing plts led to lower HSR, ADP and collagen-induced aggregation, percentage of cells expressing GP1b alpha, GPIIb and basal binding of PAC-1 compared to levels measured in fresh plts (p<0.05, all), and the authors report a reduction in plt swirl and loss of the PAC-1 response to ADP in thawed plts compared to fresh specimens. However, freezing increased the percentage of plts positive for CD62P, CD63, and annexin-V, and increased levels of CD62P, PF4, PDGF-AB, TGF-beta, beta-TG, and EGF. Levels of CD62P, PF4, PDGF-AB, TGF-beta, and EGF depended on storage solution and post-thaw storage duration at room temperature.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • Frozen
   • None (Fresh)

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry
   Cell count/volume	Hematology/ auto analyzer
   Protein	Flow cytometry
   Protein	ELISA
   Small molecule	Clinical chemistry/auto analyzer
   Morphology	Macroscopic observation
   Morphology	Flow cytometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Time at room temperature	0 h 6 h 24 h
   Storage	Freeze/thaw cycling	0 cycles 1 cycle
   Storage	Short-term storage solution	Plasma PAS-G SSP+
   Biospecimen Preservation	Type of fixation/preservation	Frozen None (fresh)
   Flow cytometry Specific	Reaction solution	ADP Collagen

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