血小板冷冻保存实验探究

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1、血小板冷冻保存实验探究作者：杨惠云田玮郭颖张瑞丽张王刚【摘要】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UDPGal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+UDPGal,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+thromb

2、osol+UDPGal组保护作用优于5%DMS0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%DMS0+thrombosol+UDPGal对血小板形态的保持效果相似，均优于5%DMS0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UDPGal对保存效果没有明显影响。【关键词】血小板；冷冻保存；thrombosol；UDPGal；CD62PExperimentalStud

3、yonCryopreservationofPlateletsAbstractThestudywaspurposedtodevelopanovelcryopreservedagent(CPA)forplatelets,toinvestigatethemorphologyofcryopreservedplateletsindifferentCPAandtheCD62Pexpressiononmembraneofplateletsafterstimulatingbythrombin,aswellastocomparetheeffectofaddingUDPGalon

4、preservedefficiencyofpreservationsolutions.Anovelcryopreservedagentconsistingof2%DMSO,thrombosolandUDPGalwasdevelopedonbasisofusinghigherconcentrationofDMS0.Themorphologyofchilledplateletswasobservedbytransmissionelectronmicroscopeandcomparedwithfreshplatelets.TheexpressionofCD62Pon

5、themembraneofplateletswasdetectedat0,1,3months.TheresuItsindicatedthatthesignificanteffectofcryopreservationonmorphologyofplateletswasfoundaccordingtopercentagesofround,dendriticandirregularshapesofcryopreservedplatelets・Theprotectiveeffectsof2%DMSO+thrombosoland2%DMSO+thrombosol+UD

6、PGalwerebetterthanthatof5%DMS0.Comparedwithfreshplatelets,theexpressionofCD62Ponplateletmembranedecreasedobviouslyaftercryopreservation,butnotobserveddifferenceatpreservationfor1monthand3months,aswellasamong3kindsofdifferentCPA.Itisconcludedthattheprotectiveeffectsof2%DMSO+thromboso

7、land2%DMSO+thrombosol+UDPGalonmorphologyofplateletsaresimilar,butbetterthanthat5%DMS0.Thereactionofcryopreservedplateletstothrombindecreases,whilethesignificantdiffereneeisnotfoundamongthese3kindsofCPA・TheadditionofUDPGaltocryopresevedagentsnotshowtheprotectiveeffectonplatelets・Keyw

8、ordsplatelet；lyophi