血小板冷冻保存的实验研究

本研究研制新型的血小板冷冻保存液,观察血小板在不同保存液冻存前、后的形态改变,以及体外受凝血酶作用下的活化状态,同时比较添加UDP-Gal对保存液保存效果的影响。在以往单一应用较高浓度的DMSO为血小板低温保存液的基础上,自行研制新型保存液：2%DMSO＋thrombosol＋UDP-Gal,在不同时间分别观察血小板形态及测定血小板膜表面糖蛋白CD62P的表达。结果显示,在圆形、树突形、不规则形态的百分比方面冷冻保存对血小板形态有显著影响,2%DMSO＋thrombosol组和2%DMSO＋thrombosol＋UDP-Gal组保护作用优于5%DM-SO组;与新鲜血小板相比,冻存后血小板表达CD62P明显下降,保存1月与3月时CD62P表达无明显差异,3种保存液组之间CD62P表达无明显差异。结论：3种冷冻保存液中2%DMSO＋thrombosol和2%DMSO＋thrombosol＋UDP-Gal对血小板形态的保持效果相似,均优于5%DMSO组。冷冻保存后血小板对凝血酶的反应下降,3种保存液组之间无明显差异。在保存液中添加UDP-Gal对保存效果没有明显影响。

Experimental Study on Cryopreservation of Platelets

The study was purposed to develop a novel cryopreserved agent (CPA) for platelets, to investigate the morphology of cryopreserved platelets in different CPA and the CD62P expression on membrane of platelets after stimulating by thrombin, as well as to compare the effect of adding UDP-Gal on preserved efficiency of preservation solutions. A novel cryopreserved agent consisting of 2% DMSO, thrombosol and UDP-Gal was developed on basis of using higher concentration of DMSO. The morphology of chilled platelets was observed by transmission electron microscope and compared with fresh platelets. The expression of CD62P on the membrane of platelets was detected at 0, l, 3 months. The results indicated that the significant effect of cryopreservation on morphology of platelets was found according to percentages of round, dendritic and irregular shapes of cryopreserved platelets. The protective effects of 2% DMSO+thrombosol and 2% DMSO+thrombosol+UDP-Gal were better than that of 5% DMSO. Compared with fresh platelets, the expression of CD62P on platelet membrane decreased obviously after cryopreservation, but not observed difference at preservation for 1 month and 3 months, as well as among 3 kinds of different CPA. It is concluded that the protective effects of 2% DMSO+thrombosol and 2% DMSO+thrombosol+UDP-Gal on morphology of platelets are similar, but better than that 5% DMSO. The reaction of cryopreserved platelets to thrombin decreases, while the significant difference is not found among these 3 kinds of CPA. The addition of UDP-Gal to cryopreseved agents not show the protective effect on platelets.