In vitro comparison of cryopreserved and liquid platelets: potential clinical implications.
Author(s): Johnson L, Reade MC, Hyland RA, Tan S, Marks DC
Publication: Transfusion, 2015, Vol. 55(4), Page 838-47
PubMed ID: 25371169 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to compare glycoprotein (GP) expression, platelet (Plt) activity, microparticle counts, and Plt function between cryopreserved and room temperature stored Plts and to determine the effects of the storage of plasma used for reconstitution of cryopreserved Plts.
Conclusion of Paper
Compared to Plts that were stored at room temperature, cryopreservation of Plts affected Plt counts, pH, expression of CD62P, CD63, GPs, basal and stimulated activated GPIIIa receptor (PAC-1), the percentage of phosphatidylserine (PS) and tissue factor (TF) positive microparticles, collagen and adenosine diphosphate (ADP) induced aggregation and the thromboelastography (TEG) parameters. The significance of these effects depended on Plt storage duration, the temperature at which the plasma used for reconstitution was deep-frozen (-80˚C, DFP) or fresh-frozen (-30˚C, FFP) and whether the FFP used for reconstitution had been refrigerated for 24 h after thawing (FFP24) or not. The supernatants of cryopreserved Plts had shorter clotting times, shorter PS and TF dependent thrombin lag times, larger PS and TF dependent thrombin peaks and higher soluble P-selectin, thromboxane and Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES) levels than supernatants of room temperature stored Plts, regardless of storage history of the plasma used for reconstitution, but significance depended on storage duration. Room temperature storage of Plts for 5 days, as opposed to 2 days, resulted in decreased Plt aggregation in response to collagen, but did not significantly affect other parameters.
Studies
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Study Purpose
The purpose of this study was to determine the effects of cryopreservation versus room temperature storage and treatment of plasma used for Plt reconstitution on Plt function, GP expression, and microparticle content. Plts obtained from 12 donors by apheresis were stored for 5 days at 22°C with shaking and were gamma irradiated on day 2. 28 non-irradiated apheresis Plt specimens were frozen in dimethyl sulfoxide (DMSO) at -80°C on day 2 post-collection and stored for 8-63 days before reconstitution in thawed 30°C plasma that had been deep-frozen at -80˚C (DFP), plasma that had been fresh frozen at -30˚C (FFP), or FFP stored at 4°C for up to 24 hours after thawing (FFP24). Plt supernatants were stored at -80°C before analysis.
Summary of Findings:
Regardless of the plasma used for reconstitution, cryopreserved Plts had higher Plt counts, pH and PS expressing microparticles, and lower GPIIb (CD41a), GPIbα (CD42b), PAC-1 expression, collagen and ADP induced aggregation, TEG time to clot (R) and clot strength (MA) than room temperature stored Plts, but significance depended on storage duration. Cryopreserved Plts reconstituted in DFP had a lower percentage of GPIIIa (CD61) and GPIX (CD42a) positive cells and a higher percentage of CD62P, CD63, and PAC-1 positive cells and TF positive microparticles than Plts stored at room temperature, but significance depended on storage duration. Cryopreserved Plts reconstituted in FFP had a higher percentage of basal PAC-1 positive cells than Plts stored at room temperature for 5 days, and those reconstituted in FFP24 had a lower percentage of GPIX (CD42a) positive Plts than Plts stored at room temperature for 2 or 5 days. The supernatants of cryopreserved Plts had shorter clotting times, shorter PS and TF dependent thrombin lag times, larger PS and TF dependent thrombin peaks, and higher soluble P-selectin, thromboxane and RANTES levels than supernatants of room temperature stored Plts, regardless of reconstitution plasma type, but significance depended on storage duration. Interleukin (IL)-27 in the supernatants was not affected by storage of Plts at room temperature or cryopreservation. Storage of Plts at room temperature for 5 days instead of 2 days resulted in decreased Plt aggregation in response to collagen, but it did not significantly affect other parameters.
Biospecimens
Preservative Types
- None (Fresh)
- Frozen
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Small molecule Clinical chemistry/auto analyzer Cell count/volume Hematology/ auto analyzer Cell count/volume Flow cytometry Protein ELISA Morphology Flow cytometry Glycoprotein Flow cytometry Morphology Thromboelastography Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 2 days
5 days
Storage Short-term storage solution DFP
FFP
FFP24
Biospecimen Preservation Type of fixation/preservation Frozen
None (fresh)
Storage Storage temperature -80˚C
-30˚C
Room temperature