Cryopreservation of buffy-coat-derived platelet concentrates in dimethyl sulfoxide and platelet additive solution.
Platelets prepared in plasma can be frozen in 6% dimethyl sulfoxide (Me(2)SO) 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% Me(2)SO. 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 24h 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 24h post-thaw, indicating an extended shelf-life may be possible. The data suggests that freezing platelets in 6% Me(2)SO and additive solution produces acceptable in vitro platelet quality.
Research and Business Development, Australian Red Cross Blood Service, 153 Clarence Street, Sydney, NSW 2000, Australia. ljohnson@arcbs.redcross.org.au
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Johnson
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Winter
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Reid
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Hartkopf-Theis
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Marks
D058621:Blood Buffy Coat / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism*; D001793:Blood Preservation / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry / Q000494:pharmacology*; D016207:Cytokines / Q000032:analysis / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000737:chemistry / Q000494:pharmacology*; D005615:Freezing; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D019344:Lactic Acid / Q000032:analysis / Q000378:metabolism; D019007:P-Selectin / Q000032:analysis / Q000378:metabolism; D010949:Plasma / Q000378:metabolism; D015539:Platelet Activation; D010974:Platelet Aggregation; D010976:Platelet Count; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000032:analysis / Q000378:metabolism; D012996:Solutions / Q000737:chemistry / Q000494:pharmacology
D003451:Cryoprotective Agents; D016207:Cytokines; D019007:P-Selectin; D019038:Platelet Glycoprotein GPIb-IX Complex; D012996:Solutions; C023592:glycocalicin; D019344:Lactic Acid; D005947:Glucose; D004121:Dimethyl Sulfoxide