Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, Maryland 20892, USA.
Transfusion. 2011 Dec;51(12):2703-8. doi: 10.1111/j.1537-2995.2011.03258.x. Epub 2011 Jul 25.
Red blood cell (RBC) preservation is essential to transfusion medicine. Many blood group reference laboratories need a method to preserve rare blood samples for serologic testing at a later date. This study offers a comparison of three common cryoprotective agents and protocols used today: bulk preservation with glycerol and droplet freezing with sucrose-dextrose (S+D) or polyvinylpyrrolidone (PVP).
Human blood from 14 volunteers was collected and frozen at set intervals over 2 weeks with PVP, S+D, or glycerol. The frozen RBCs were later thawed and the percentage of surviving RBCs was determined. Detailed protocols and an instructional video are supplied.
Over a 2-week period, RBCs preserved with glycerol and thawed with a widely used protocol showed a recovery of 41 ± 16% (mean ± standard deviation) while those thawed with a modified glycerol protocol showed a recovery of 76 ± 8%. RBCs preserved by droplet freezing with S+D showed a recovery of 56 ± 11% while those preserved by droplet freezing with PVP showed a recovery of 85 ± 6%. Recovery values were similar with ethylenediaminetetraacetic acid or heparin anticoagulants, differing freezing rates, and varying droplet volumes.
Droplet freezing with PVP offered the greatest recovery. While bulk freezing with glycerol can also be effective, droplet freezing may be a more convenient method overall. It requires less effort to thaw, needs much less storage room, and allows blood group laboratories to be frugal with thawing rare samples.
红细胞(RBC)的保存对于输血医学至关重要。许多血型参考实验室需要一种方法来保存稀有血液样本,以便日后进行血清学检测。本研究比较了三种常用的冷冻保护剂和目前使用的三种方案:甘油的批量保存、蔗糖-葡萄糖(S+D)或聚乙烯吡咯烷酮(PVP)的液滴冷冻。
从 14 名志愿者采集人血,在 2 周内用 PVP、S+D 或甘油分阶段冷冻。随后解冻冷冻的 RBC,确定存活 RBC 的百分比。提供了详细的方案和教学视频。
在 2 周的时间内,用甘油保存并采用广泛使用的方案解冻的 RBC 恢复率为 41±16%(平均值±标准差),而用改良甘油方案解冻的 RBC 恢复率为 76±8%。用 S+D 液滴冷冻保存的 RBC 恢复率为 56±11%,而用 PVP 液滴冷冻保存的 RBC 恢复率为 85±6%。用乙二胺四乙酸或肝素抗凝剂、不同的冷冻速率和不同的液滴体积,回收值相似。
用 PVP 液滴冷冻可获得最大的恢复率。虽然用甘油批量冷冻也可以有效,但液滴冷冻总体上可能是一种更方便的方法。它需要更少的解冻努力,需要更少的存储空间,并允许血型实验室在解冻稀有样本时更加节约。