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Cryopreservation of buffy-coat-derived platelet concentrates in dimethyl sulfoxide and platelet additive solution
Authors:Johnson L N  Winter K M  Reid S  Hartkopf-Theis T  Marks D C
Affiliation:Research and Business Development, Australian Red Cross Blood Service, Sydney, Australia
Abstract:Platelets prepared in plasma can be frozen in 6% dimethyl sulfoxide (Me2SO) and stored for extended periods at −80 °C. The aim of this study was to reduce the plasma present in the cryopreserved product, by substituting plasma with platelet additive solution (PAS; SSP+), whilst maintaining in vitro platelet quality. Buffy coat-derived pooled leukoreduced platelet concentrates were frozen in a mixture of SSP+, plasma and 6% Me2SO. The platelets were concentrated, to avoid post-thaw washing, and frozen at −80 °C. The cryopreserved platelet units (n = 9) were rapidly thawed at 37 °C, reconstituted in 50% SSP+/plasma and stored at 22 °C. Platelet recovery and quality were examined 1 and 24 h post-thaw and compared to the pre-freeze samples. Upon thawing, platelet recovery ranged from 60% to 80%. However, there were differences between frozen and liquid-stored platelets, including a reduction in aggregation in response to ADP and collagen; increased CD62P expression; decreased viability; increased apoptosis and some loss of mitochondrial membrane integrity. Some recovery of these parameters was detected at 24 h post-thaw, indicating an extended shelf-life may be possible. The data suggests that freezing platelets in 6% Me2SO and additive solution produces acceptable in vitro platelet quality.
Keywords:Abbreviations: Me2SO, dimethyl sulfoxide   PAS, platelet additive solution   HSR, hypotonic shock response   ADP, adenosine diphosphate   PS, phosphatidylserine   Δψ, mitochondrial membrane potential   TRALI, transfusion related acute lung injury
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