Langer R, Albrecht R, Hempel K, Krug S, Sputtek A, Steigerwald R, Trenkel K, Henrich H A
Abteilung Experimentelle Chirurgie, Chirurgische Universitätsklinik, Würzburg.
Infusionsther Transfusionsmed. 1994 Dec;21(6):393-400.
Cryopreservation of erythrocytes using hydroxyethyl starch (HES) as cryoprotecting additive could result in a nearly unlimited storage stability of preserved red cells. In addition, it would allow its immediate use for transfusion. In order to assess the therapeutic efficacy of erythrocytes cryopreserved with HES, their 24-hour post-transfusion survival and long-term survival was evaluated.
The experiments were carried out with dog erythrocytes as an animal model for human erythrocytes. To each of 6 German shepherd dogs a 15-ml sample of erythrocyte suspension, labeled with 51Cr (25 microCi) after thawing, was autologously injected. Caused by hemolysis 29% of the formerly cryopreserved erythrocytes have not been labeled. To each of 6 control animals 15 ml of a suspension of freshly drawn and 51Cr-labeled erythrocytes was injected. The 51Cr radioactivity in later taken blood samples was a measure for the number of injected erythrocytes having remained in the circulation until the moment of blood withdrawal. The effect of cryopreservation was assessed by comparison of the test group with the control group.
In both groups 30% of the applied cells left the circulation within 30 min. This was effected by pharmacological enlargement of the dogs' spleen and not by hemolysis of the erythrocytes. After the first 24 h all of the cryopreserved labeled erythrocytes had survived to the same amount (> 95%) as the labeled fresh red cells. Between 12 h and 20 days after injection, in both groups the 51Cr activity decreased exponentially by 4.8 and 4.5%/d. This difference was not significant. The area under the curve amounted to 1253 and 1257% d, respectively.
There exists a subpopulation of red cells that is destroyed by freezing stress. As a result the freed stroma would be a serious transfusion risk. All erythrocytes having survived the cryopreservation procedure resemble the fresh erythrocytes with regard to the in-vivo survival; their therapeutic efficacy is not impaired. In the context of in-vitro results with human erythrocytes it can be expected that at the present developmental state of the cryopreservation procedure at least 93% of the human erythrocytes cryopreserved with HES have a normal 24-hour and long-term post-transfusion survival.
使用羟乙基淀粉(HES)作为冷冻保护添加剂对红细胞进行冷冻保存可使保存的红细胞具有几乎无限的储存稳定性。此外,这还能使其可立即用于输血。为了评估用HES冷冻保存的红细胞的治疗效果,对其输血后24小时的存活率和长期存活率进行了评估。
以犬红细胞作为人类红细胞的动物模型进行实验。给6只德国牧羊犬每只自体注射15毫升解冻后用51铬(25微居里)标记的红细胞悬液。由于溶血,之前冷冻保存的红细胞中有29%未被标记。给6只对照动物每只注射15毫升新采集的并用51铬标记的红细胞悬液。后来采集的血样中的51铬放射性是对直至采血时仍留在循环中的注射红细胞数量的一种衡量。通过将试验组与对照组进行比较来评估冷冻保存的效果。
在两组中,30%的注入细胞在30分钟内离开循环。这是通过药物扩大犬的脾脏实现的,而非红细胞溶血所致。在最初的24小时后,所有冷冻保存的标记红细胞存活数量与标记的新鲜红细胞相同(>95%)。在注射后12小时至20天之间,两组中的51铬活性均以每天4.8%和4.5%的速度呈指数下降。这种差异不显著。曲线下面积分别为1253和1257%·天。
存在一部分红细胞亚群会因冷冻应激而被破坏。结果释放出的基质会带来严重的输血风险。所有在冷冻保存过程中存活下来的红细胞在体内存活率方面与新鲜红细胞相似;其治疗效果未受损害。结合对人类红细胞的体外实验结果,可以预期,在目前冷冻保存方法的发展状态下,至少93%用HES冷冻保存的人类红细胞具有正常的输血后24小时和长期存活率。