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二甲基亚砜和血小板添加剂溶液中来源于浓缩血小板白细胞层的血小板的冷冻保存。

Cryopreservation of buffy-coat-derived platelet concentrates in dimethyl sulfoxide and platelet additive solution.

机构信息

Research and Business Development, Australian Red Cross Blood Service, 153 Clarence Street, Sydney, NSW 2000, Australia.

出版信息

Cryobiology. 2011 Apr;62(2):100-6. doi: 10.1016/j.cryobiol.2011.01.003. Epub 2011 Jan 15.

Abstract

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.

摘要

血小板可在含有 6%二甲基亚砜(Me2SO)的血浆中制备,然后在-80°C 下冷冻保存延长时间。本研究的目的是通过用血小板添加剂溶液(PAS;SSP+)替代血浆来减少冷冻产品中的血浆,同时保持体外血小板质量。从富白细胞的浓缩血小板中提取的富含血小板的白细胞减少物在 SSP+、血浆和 6%Me2SO 的混合物中冷冻。浓缩血小板,以避免解冻后洗涤,并在-80°C 下冷冻。将冷冻的血小板单位(n=9)在 37°C 下快速解冻,在 50%SSP+/血浆中重构并在 22°C 下储存。在解冻后 1 和 24 小时检查血小板回收率和质量,并与预冻样本进行比较。解冻后,血小板回收率在 60%至 80%之间变化。然而,冷冻和液体储存的血小板之间存在差异,包括对 ADP 和胶原的聚集反应减少;CD62P 表达增加;活力降低;凋亡增加和一些线粒体膜完整性丧失。在解冻后 24 小时检测到这些参数的一些恢复,表明可能具有延长的货架寿命。数据表明,在 6%Me2SO 和添加剂溶液中冷冻血小板可产生可接受的体外血小板质量。

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