Calcium chelation: a novel approach to reduce cryopreservation-induced damage to frozen platelets.
Transfusion
; 60(7): 1552-1563, 2020 07.
Article
em En
| MEDLINE
| ID: mdl-32319689
ABSTRACT
BACKGROUND:
Cryopreserved platelets are phenotypically and functionally different to conventionally stored platelets. Calcium may be released from internal stores during the freeze-thaw process, initiating signaling events which lead to these alterations. It was hypothesized that the addition of a calcium chelator prior to cryopreservation may mitigate some of these changes.METHODS:
Buffy coat-derived platelets that had been pooled and split were tested fresh and following cryopreservation (n = 8 per group). Platelets were cryopreserved using 5%-6% dimethylsulfoxide (DMSO) or were supplemented with increasing concentrations of the internal calcium chelator, BAPTA-AM (100 µM, 200 µM, or 400 µM), prior to storage at -80°C.RESULTS:
Supplementation of platelets with BAPTA-AM prior to freezing improved platelet recovery in a dose response manner (400 µM 84 ± 2%) compared to standard DMSO cryopreserved platelets (70 ± 4%). There was a loss of GPIbα, GPVI, and GPIIb/IIIa receptors on platelets following cryopreservation, which was rescued when platelets were supplemented with BAPTA-AM (400 µM p < 0.0001 for all). Platelet activation markers, such as phosphatidylserine and P-selectin, were externalized on platelets following cryopreservation. However, the addition of BAPTA-AM significantly reduced the increase of these activation markers on cryopreserved platelets (400 µM p < 0.0001 for both). Both cryopreserved platelet groups exhibited similar functionality as assessed by thromboelastography, forming clots at a faster rate than fresh platelets.CONCLUSIONS:
This study demonstrates that calcium plays a crucial role in mediating cryopreservation-induced damage to frozen platelets. The addition of the calcium chelator, BAPTA-AM, prior to cryopreservation reduces this damage.
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Base de dados:
MEDLINE
Assunto principal:
Plaquetas
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Preservação de Sangue
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Criopreservação
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Dimetil Sulfóxido
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Cálcio
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Ácido Egtázico
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Quelantes de Cálcio
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article