Calcium chelation: a novel approach to reduce cryopreservation-induced damage to frozen platelets.

Waters, L; Padula, MP; Marks, DC; Johnson, L

Calcium chelation: a novel approach to reduce cryopreservation-induced damage to frozen platelets.

Waters, L

Padula, MP Marks, DC Johnson, L

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Publisher:: WILEY
Publication Type:: Journal Article
Citation:: Transfusion, 2020, 60, (7), pp. 1552-1563
Issue Date:: 2020-07

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Field	Value	Language
dc.contributor.author	Waters, L https://orcid.org/0000-0002-4740-055X
dc.contributor.author	Padula, MP
dc.contributor.author	Marks, DC
dc.contributor.author	Johnson, L https://orcid.org/0000-0003-3303-4437
dc.date.accessioned	2020-11-05T05:28:15Z
dc.date.available	2021-08-12T19:01:06Z
dc.date.issued	2020-07
dc.identifier.citation	Transfusion, 2020, 60, (7), pp. 1552-1563
dc.identifier.issn	0041-1132
dc.identifier.issn	1537-2995
dc.identifier.uri	http://hdl.handle.net/10453/143775
dc.description.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.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	WILEY
dc.relation.ispartof	Transfusion
dc.relation.isbasedon	10.1111/trf.15799
dc.rights	info:eu-repo/semantics/embargoedAccess
dc.rights	Wiley "This is the peer reviewed version of the following article: Waters, Lauren; Padula, Matthew P.; Marks, Denese C.; Johnson, Lacey. Calcium chelation: a novel approach to reduce cryopreservation-induced damage to frozen platelets. (2020) Transfusion; doi: 10.1111/trf.15799 which has been published in final form at 10.1111/jan.14128 ”. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving."
dc.subject	1102 Cardiorespiratory Medicine and Haematology, 1103 Clinical Sciences, 1107 Immunology
dc.subject.classification	Cardiovascular System & Hematology
dc.title	Calcium chelation: a novel approach to reduce cryopreservation-induced damage to frozen platelets.
dc.type	Journal Article
utslib.citation.volume	60
utslib.location.activity	United States
utslib.for	1102 Cardiorespiratory Medicine and Haematology
utslib.for	1103 Clinical Sciences
utslib.for	1107 Immunology
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney
utslib.copyright.status	open_access	*
pubs.consider-herdc	false
dc.date.updated	2020-11-05T05:28:09Z
pubs.issue	7
pubs.publication-status	Published
pubs.volume	60
utslib.citation.issue	7

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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http://hdl.handle.net/10453/143775