Dijkstra-Tiekstra M J, de Korte D, Pietersz R N I, Reesink H W, van der Meer P F, Verhoeven A J
Sanquin Blood Bank north west Region, Amsterdam, The Netherlands.
Vox Sang. 2003 Nov;85(4):276-82. doi: 10.1111/j.0042-9007.2003.00360.x.
Leucoreduced platelet concentrates (LR-PCs) can be stored at 20-24 degrees C for 5-7 days. When LR-PCs are cryopreserved they can be stored for several years. For cryopreservation to become applicable in blood-bank practice, an off-the-shelf cryoprotectant is needed that can be added to the LR-PC in a sterile manner. For this, we varied the composition of the cryopreservation medium and studied various parameters of cryopreserved LR-PCs for up to 24 h after thawing at room temperature.
LR-PCs in plasma or Composol were concentrated and divided into 2 units. To each unit, an equal part of 10% dimethylsulfoxide (DMSO) in plasma, Composol with or without 5% albumin, or GPO (pasteurized plasma-protein solution) was added. Freezing occurred at 1 degrees C/min and LR-PCs were placed in the vapour phase of nitrogen. LR-PCs were thawed at 37 degrees C and stored at room temperature. LR-PCs were tested for morphology, platelet recovery, swirling effect, and activation antigens at various time-points thereafter.
LR-PCs in 100%, 65% and 50% plasma supplemented with Composol showed good morphology scores (>250), limited CD62P expression (<35%), low CD63 expression (<20%) and a swirling effect of about 2, at 24 h after thawing. At the same time-point, platelet recovery was >80% under all conditions and CD42b expression varied between 70 and 85%. Results of LR-PCs in 15% plasma and Composol, with or without plasma substitutes, were not acceptable at 24 h after thawing, i.e. the morphology score was <200 and the CD62P expression was >40%.
A minimum of 50% plasma in the cryopreserved LR-PC is necessary to maintain an acceptable in vitro quality of platelets up to 24 h after thawing. Composol is a good candidate for using to prepare an off-the-shelf cryoprotectant.
白细胞滤除血小板浓缩物(LR-PCs)可在20-24℃下储存5-7天。当LR-PCs被冷冻保存时,它们可以储存数年。为了使冷冻保存适用于血库实践,需要一种现成的冷冻保护剂,它可以以无菌方式添加到LR-PC中。为此,我们改变了冷冻保存介质的成分,并研究了在室温下解冻后长达24小时的冷冻保存LR-PCs的各种参数。
将血浆或Composol中的LR-PCs浓缩并分成2个单位。向每个单位中加入等量的血浆中10%二甲基亚砜(DMSO)、含或不含5%白蛋白的Composol或GPO(巴氏消毒血浆蛋白溶液)。以1℃/分钟的速度冷冻,将LR-PCs置于氮气的气相中。LR-PCs在37℃下解冻并在室温下储存。此后在不同时间点对LR-PCs进行形态学、血小板回收率、漩涡效应和活化抗原检测。
在解冻后24小时,补充Composol的100%、65%和50%血浆中的LR-PCs显示出良好的形态学评分(>250)、有限的CD62P表达(<35%)、低CD63表达(<20%)和约为2的漩涡效应。在同一时间点,所有条件下血小板回收率均>80%,CD42b表达在70%至85%之间变化。在解冻后24小时,含或不含血浆替代品的15%血浆和Composol中的LR-PCs结果不可接受,即形态学评分<200且CD62P表达>40%。
冷冻保存的LR-PC中至少需要50%的血浆,以在解冻后长达24小时维持可接受的血小板体外质量。Composol是用于制备现成冷冻保护剂的良好候选物。