Research and Business Development, Australian Red Cross Blood Service, 153 Clarence Street, Sydney, NSW 2000, Australia.
Cryobiology. 2011 Apr;62(2):100-6. doi: 10.1016/j.cryobiol.2011.01.003. Epub 2011 Jan 15.
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.
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。