期刊
TRANSFUSION
卷 61, 期 9, 页码 2549-2555出版社
WILEY
DOI: 10.1111/trf.16542
关键词
cryopreservation; mass spectrometry; platelets; protein synthesis; proteomics
类别
资金
- Australian Red Cross Lifeblood
- Canadian Blood Services
- Health Canada
Cryo-preserved platelets are capable of synthesizing proteins upon reconstitution and storage, with a subset of these proteins observed in platelet microvesicles (PMVs). This discovery suggests a possible role in vesicle encapsulation, providing insights into protein synthesis and packaging regulation in cryo-PLTs.
Background Cryopreservation of platelets (PLTs) could allow extension of their shelf-life to years, compared to days for liquid stored platelets. Due to their greater hemostatic effect, reconstituted cryopreserved platelets (cryo-PLTs) would be able to support bleeding emergencies. Since protein synthesis has been linked to PLT functions, such as clot formation and immune responses, the translational capacity of reconstituted cryo-PLTs was assessed upon thawing and short-term storage. Methods/materials Platelets were frozen at -80 degrees C with 5-6% DMSO. Upon thawing, they were reconstituted in plasma and then aliquoted (12 ml) into mini-bags and assessed over 24 h of storage at RT. One series served as control; the second and third series were spiked with either 300 mu M puromycin (Pm) or 227 nM biotin-labeled Pm. Samples were tested for in vitro quality and PLT microvesicle enumeration by flow cytometry. Protein synthesis in cryo-PLTs was assessed using a modified method based on puromycin-associated nascent chain proteomics. Results In vitro parameters of reconstituted and subsequently stored platelets were consistent with previously published results. Mass-spectrometry analyses identified that 22 proteins were synthesized in PLTs and 13 of those were observed in platelet microvesicles (PMVs). Conclusion Cryo-PLTs can synthesize proteins upon reconstitution and storage. Discovery of a subset of these proteins in the PMV suggests a role in vesicle encapsulation, possibly in a selective manner. This observation provides novel insights into the capacity for protein synthesis in cryo-PLTs and the potential regulation of protein packaging into PMV.
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