Identification of platelet subpopulations in cryopreserved platelet components using multi-colour imaging flow cytometry.

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

Identification of platelet subpopulations in cryopreserved platelet components using multi-colour imaging flow cytometry.

Johnson, L Lei, P Waters, L Padula, MP Marks, DC

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Publisher:: Springer Nature
Publication Type:: Journal Article
Citation:: Sci Rep, 2023, 13, (1), pp. 1221
Issue Date:: 2023-01-21

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Field	Value	Language
dc.contributor.author	Johnson, L
dc.contributor.author	Lei, P
dc.contributor.author	Waters, L
dc.contributor.author	Padula, MP
dc.contributor.author	Marks, DC
dc.date.accessioned	2023-02-15T06:08:38Z
dc.date.available	2023-01-17
dc.date.available	2023-02-15T06:08:38Z
dc.date.issued	2023-01-21
dc.identifier.citation	Sci Rep, 2023, 13, (1), pp. 1221
dc.identifier.issn	2045-2322
dc.identifier.issn	2045-2322
dc.identifier.uri	http://hdl.handle.net/10453/166159
dc.description.abstract	Cryopreservation of platelets, at - 80 °C with 5-6% DMSO, results in externalisation of phosphatidylserine and the formation of extracellular vesicles (EVs), which may mediate their procoagulant function. The phenotypic features of procoagulant platelets overlap with other platelet subpopulations. The aim of this study was to define the phenotype of in vitro generated platelet subpopulations, and subsequently identify the subpopulations present in cryopreserved components. Fresh platelet components (n = 6 in each group) were either unstimulated as a source of resting platelets; or stimulated with thrombin and collagen to generate a mixture of aggregatory and procoagulant platelets; calcium ionophore (A23187) to generate procoagulant platelets; or ABT-737 to generate apoptotic platelets. Platelet components (n = 6) were cryopreserved with DMSO, thawed and resuspended in a unit of thawed plasma. Multi-colour panels of fluorescent antibodies and dyes were used to identify the features of subpopulations by imaging flow cytometry. A combination of annexin-V (AnnV), CD42b, and either PAC1 or CD62P was able to distinguish the four subpopulations. Cryopreserved platelets contained procoagulant platelets (AnnV+/PAC1-/CD42b+/CD62P+) and a novel population (AnnV+/PAC1-/CD42b+/CD62P-) that did not align with the phenotype of aggregatory (AnnV-/PAC1+/CD42b+/CD62P+) or apoptotic (AnnV+/PAC1-/CD42b-/CD62P-) subpopulations. These data suggests that the enhanced haemostatic potential of cryopreserved platelets may be due to the cryo-induced development of procoagulant platelets, and that additional subpopulations may exist.
dc.format	Electronic
dc.language	eng
dc.publisher	Springer Nature
dc.relation.ispartof	Sci Rep
dc.relation.isbasedon	10.1038/s41598-023-28352-2
dc.rights	info:eu-repo/semantics/openAccess
dc.subject.mesh	Dimethyl Sulfoxide
dc.subject.mesh	Flow Cytometry
dc.subject.mesh	Color
dc.subject.mesh	Blood Platelets
dc.subject.mesh	Cryopreservation
dc.subject.mesh	Platelet Activation
dc.subject.mesh	Blood Platelets
dc.subject.mesh	Dimethyl Sulfoxide
dc.subject.mesh	Cryopreservation
dc.subject.mesh	Flow Cytometry
dc.subject.mesh	Platelet Activation
dc.subject.mesh	Color
dc.subject.mesh	Dimethyl Sulfoxide
dc.subject.mesh	Flow Cytometry
dc.subject.mesh	Color
dc.subject.mesh	Blood Platelets
dc.subject.mesh	Cryopreservation
dc.subject.mesh	Platelet Activation
dc.title	Identification of platelet subpopulations in cryopreserved platelet components using multi-colour imaging flow cytometry.
dc.type	Journal Article
utslib.citation.volume	13
utslib.location.activity	England
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	open_access	*
dc.date.updated	2023-02-15T06:08:32Z
pubs.issue	1
pubs.publication-status	Published online
pubs.volume	13
utslib.citation.issue	1

Abstract:

Cryopreservation of platelets, at - 80 °C with 5-6% DMSO, results in externalisation of phosphatidylserine and the formation of extracellular vesicles (EVs), which may mediate their procoagulant function. The phenotypic features of procoagulant platelets overlap with other platelet subpopulations. The aim of this study was to define the phenotype of in vitro generated platelet subpopulations, and subsequently identify the subpopulations present in cryopreserved components. Fresh platelet components (n = 6 in each group) were either unstimulated as a source of resting platelets; or stimulated with thrombin and collagen to generate a mixture of aggregatory and procoagulant platelets; calcium ionophore (A23187) to generate procoagulant platelets; or ABT-737 to generate apoptotic platelets. Platelet components (n = 6) were cryopreserved with DMSO, thawed and resuspended in a unit of thawed plasma. Multi-colour panels of fluorescent antibodies and dyes were used to identify the features of subpopulations by imaging flow cytometry. A combination of annexin-V (AnnV), CD42b, and either PAC1 or CD62P was able to distinguish the four subpopulations. Cryopreserved platelets contained procoagulant platelets (AnnV+/PAC1-/CD42b+/CD62P+) and a novel population (AnnV+/PAC1-/CD42b+/CD62P-) that did not align with the phenotype of aggregatory (AnnV-/PAC1+/CD42b+/CD62P+) or apoptotic (AnnV+/PAC1-/CD42b-/CD62P-) subpopulations. These data suggests that the enhanced haemostatic potential of cryopreserved platelets may be due to the cryo-induced development of procoagulant platelets, and that additional subpopulations may exist.

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http://hdl.handle.net/10453/166159