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Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols

BACKGROUND: Cryopreserved platelets show a reduced recovery and viability after freezing and thawing including several ultrastructural and phenotypic deteriorations compared with liquid‐stored platelets. It is suggested that using Controlled‐Rate Freezing (CRF) can reduce variability and optimize th...

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Autores principales: Tynngård, Nahreen, Bell, Alice, Gryfelt, Gunilla, Cvetkovic, Stefan, Wikman, Agneta, Uhlin, Michael, Sandgren, Per
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7898315/
https://www.ncbi.nlm.nih.gov/pubmed/33345368
http://dx.doi.org/10.1111/trf.16227
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author Tynngård, Nahreen
Bell, Alice
Gryfelt, Gunilla
Cvetkovic, Stefan
Wikman, Agneta
Uhlin, Michael
Sandgren, Per
author_facet Tynngård, Nahreen
Bell, Alice
Gryfelt, Gunilla
Cvetkovic, Stefan
Wikman, Agneta
Uhlin, Michael
Sandgren, Per
author_sort Tynngård, Nahreen
collection PubMed
description BACKGROUND: Cryopreserved platelets show a reduced recovery and viability after freezing and thawing including several ultrastructural and phenotypic deteriorations compared with liquid‐stored platelets. It is suggested that using Controlled‐Rate Freezing (CRF) can reduce variability and optimize the functionality profile for cells. The objective of the study is to compare cellular, metabolic, phenotypic and functional effects on platelets after cryopreservation using different freezing rate protocols. STUDY DESIGN AND METHODS: To evaluate the possible effects of different freezing rate protocols a two‐experimental study comparing diverse combinations was tested with a pool and split design. Uncontrolled freezing of platelets in materials with different thermal conductivity (metal vs cardboard) was evaluated in experiment 1. Experiment 2 evaluated uncontrolled vs a controlled‐rate freezing protocol in metal boxes. All variables were assessed pre and post cryopreservation. RESULTS: Directly after thawing, no major differences in platelet recovery, LDH, ATP, Δψ, CD62P, CD42b, platelet endothelial cell adhesion molecule and sCD40L were seen between units frozen with different thermal conductivity for temperature. In contrast, we observed signs of increased activation after freezing using the CRF protocol, reflected by increased cell surface expression of CD62P, PAC‐1 binding and increased concentration of LDH. Agonist induced expression of a conformational epitope on the GPIIb/IIIa complex and contribution to blood coagulation in an experimental rotational thromboelastometry setup were not statistically different between the groups. CONCLUSION: The use of a uncontrolled freezing rate protocol is feasible, creating a platelet product comparable to using a controlled rate freezing equipment during cryopreservation of platelets.
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spelling pubmed-78983152021-03-03 Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols Tynngård, Nahreen Bell, Alice Gryfelt, Gunilla Cvetkovic, Stefan Wikman, Agneta Uhlin, Michael Sandgren, Per Transfusion Blood Components BACKGROUND: Cryopreserved platelets show a reduced recovery and viability after freezing and thawing including several ultrastructural and phenotypic deteriorations compared with liquid‐stored platelets. It is suggested that using Controlled‐Rate Freezing (CRF) can reduce variability and optimize the functionality profile for cells. The objective of the study is to compare cellular, metabolic, phenotypic and functional effects on platelets after cryopreservation using different freezing rate protocols. STUDY DESIGN AND METHODS: To evaluate the possible effects of different freezing rate protocols a two‐experimental study comparing diverse combinations was tested with a pool and split design. Uncontrolled freezing of platelets in materials with different thermal conductivity (metal vs cardboard) was evaluated in experiment 1. Experiment 2 evaluated uncontrolled vs a controlled‐rate freezing protocol in metal boxes. All variables were assessed pre and post cryopreservation. RESULTS: Directly after thawing, no major differences in platelet recovery, LDH, ATP, Δψ, CD62P, CD42b, platelet endothelial cell adhesion molecule and sCD40L were seen between units frozen with different thermal conductivity for temperature. In contrast, we observed signs of increased activation after freezing using the CRF protocol, reflected by increased cell surface expression of CD62P, PAC‐1 binding and increased concentration of LDH. Agonist induced expression of a conformational epitope on the GPIIb/IIIa complex and contribution to blood coagulation in an experimental rotational thromboelastometry setup were not statistically different between the groups. CONCLUSION: The use of a uncontrolled freezing rate protocol is feasible, creating a platelet product comparable to using a controlled rate freezing equipment during cryopreservation of platelets. John Wiley & Sons, Inc. 2020-12-21 2021-02 /pmc/articles/PMC7898315/ /pubmed/33345368 http://dx.doi.org/10.1111/trf.16227 Text en © 2020 The Authors. Transfusion published by Wiley Periodicals LLC. on behalf of AABB. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Blood Components
Tynngård, Nahreen
Bell, Alice
Gryfelt, Gunilla
Cvetkovic, Stefan
Wikman, Agneta
Uhlin, Michael
Sandgren, Per
Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols
title Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols
title_full Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols
title_fullStr Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols
title_full_unstemmed Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols
title_short Cryopreservation of buffy coat derived platelets: Paired in vitro characterization using uncontrolled versus controlled freezing rate protocols
title_sort cryopreservation of buffy coat derived platelets: paired in vitro characterization using uncontrolled versus controlled freezing rate protocols
topic Blood Components
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7898315/
https://www.ncbi.nlm.nih.gov/pubmed/33345368
http://dx.doi.org/10.1111/trf.16227
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