不同型号血细胞分离机单采血小板的研究

目的观察采前血小板(PLT)计数与循环全血处理量的关系,比较不同型号血细胞分离机机采血小板的收集效率。方法随机选择207名献血员,用H aem onetics公司的三种不同型号血细胞分离机单采献血者的血小板,用血细胞计数仪计数机采血小板(PC)中的血细胞,计算三种机型的收集效率。结果发现采前血小板计数与全血处理量有密切关系,M CS 的机采血小板的白细胞污染比M CS2P、M CS3P的白细胞污染要少,统计有显著性差异。红细胞污染、血小板收集以及收集率上,三种机型无显著性差异。结论采前血小板计数与全血处理量呈负相关,在白细胞污染上M CS 机采血小板明显要好于M CS2P、M CS3P。     

机采血小板悬液在保存过程中凝血因子Ⅷ、Ⅸ生物活性的变化

为了研究机采血小板悬液22℃振荡保存不同时间的凝血因子Ⅷ、Ⅸ生物活性的变化,采用SYSMEXCA-1500型全自动血凝仪对用CS-3000plus血细胞分离机采集的18份血小板悬液于22℃振荡保存条件下,测定0、12、24、48、72、96、120小时7个时间段的FⅧ∶C和FⅨ∶C的活性。结果表明:机采血小板FⅧ∶C在0时活性为(100.51±44.02)%,保存12-120小时,其活性衰减了10%-40%;FⅨ∶C在0时活性为(120.93±20.50)%,保存24-120小时,其活性衰减了10%-35%。结论:机采血小板悬液于22℃振荡保存时凝血因子Ⅷ、Ⅸ仍保持有较高的生物学活性。     

采集后6小时与72小时制备的冰冻单采血小板质量研究

目的通过比较采集后6h和72h制备的冰冻单采血小板制品的多项质量参数,为冰冻单采血小板的制备标准提供依据。方法以美国产Trima血细胞分离机配套全密闭7d保存袋采集的单采血小板,分别于采集后6h和72h制备为冰冻血小板制品,1周后复苏留样,分别检测血小板计数（PLT,MPV,PCT,PDW）、血小板粘附性、血小板P选择蛋白、PF3A、PF4、pH值、血块收缩试验（血浆法）。结果采集后6h和72h制备的冰冻单采血小板,两者相比差异无统计学意义（t〈0．01,P〉0．05）。单采血小板在不同保存条件下,PF3A均保持了良好的PF3活性,单采血小板在常规或冰冻保存条件下,PF4及P选择蛋白的表达均明显增加,采集后6h和72h制备的冰冻单采血小板相比,差异无统计学意义（t〈0．01,P〉0．05）;单采血小板在常规保存3d内,粘附功能和血块收缩试验没有明显变化,而冰冻保存的单采血小板这两项指标呈明显减退现象,血小板的活力下降明显。结论以美国产Trima血细胞分离机配套全密闭7d保存袋采集的单采血小板,分别于采集后6h和72h制备的冰冻血小板制品,两者的体外指标分析结果相比差异无显著性,且都能有效的改善和控制急性大出血患者的出血倾向,用于抢救急性大出血患者疗效是可靠的。     

不同常规保存血小板冰冻保存效果研究

目的了解常规保存不同时间的机采血小板冰冻前后质量指标变化及输注疗效，探讨血小板冰冻处理前常规保存调控的最佳方案。方法对120袋机采血小板随机分成6组，在（22±2）℃平床振荡条件下，分别保存0、1、2、3、4、5d然后制备冰冻血小板，并对血小板冰冻前与复温后分别计数血小板，检测pH值，跟踪调查输注冰冻血小板的患者，计算回收率。结果有效期内（22±2）℃振荡保存的血小板产品中血小板计数无显著性差异，pH值下降明显；冰冻前后血小板计数有显著性差异，pH值无差异。保存3d内的血小板冰冻后血小板的输注回收率无差异，与保存4d、5d的血小板冰冻后的回收率有显著差异。结论（22±2）℃振荡保存3d内的血小板可以制备冰冻血小板，保存4-5d的血小板可以输注但不宜制备冰冻血小板。     

机采血小板保存期内血小板聚集功能和可溶性P-选择蛋白的变化

本研究探讨保存期内的机采血小板的聚集功能和可溶性P-选择蛋白含量的改变。收集20份机采血小板样品，分别在保存期第1天（0—24小时）、第2天（24—48小时）、第3天（48—72小时）、第4天（72—96小时）和第5天（96—120小时）检测血小板的聚集功能和可溶性P-选择蛋白含量。结果表明：以ADP作为诱导剂，保存期内机采血小板的血小板聚集功能降低明显，与第1天组比，组间差异均有统计学意义（P均〈0．01），第4天组的血小板最大聚集率≤3％；血浆可溶性P-选择素含量则随着保存时间的延长逐渐升高，与第1天组比，组间差异均有统计学意义（p均〈0．05）。结论：机采血小板在保存期内存在持续活化，且聚集功能下降明显，从第4天开始几乎已完全丧失对ADP的致聚反应，提示机采血小板的体外保存损伤应该引起高度重视。     

常温保存期血小板数量及功能的变化

目的探讨保存期机采血小板数量及功能的变化。方法血细胞分离机采集血小板8人份,22℃振荡保存,分别在保存0、1、3和5d,在100级超净台内取样10ml,进行血小板计数、pH值、乳酸脱氢酶(LDH)、血小板凋亡数及CD62P表达的检测。结果在0、1、3和5d的Plt没有显著性差异;pH值比较,有显著性差异;0d与3d和5d相比,LDH有显著性差异;0d与第1、3、5d相比,血小板凋亡数有显著性差异;0d与1d、5d相比,CD62P表达有显著性差异。结论在血小板保存期,随着保存期的延长,机采血小板的pH值下降;LDH水平逐步增高;血小板凋亡数逐渐增高;CD62P的表达也逐渐增高,激活的血小板数增多。     

25Gy γ射线辐照对手工富集人血小板CD62p表达的影响

本研究观察25Gyγ射线照射对22℃保存条件下的手工富集血小板悬液不同保存期CD62p、血小板计数及平均血小板体积(MPV)的影响。将富浆法分离的16袋血小板,每袋平均分为两份,其中1袋经25Gy137铯γ射线辐照,另1份不辐照,然后按美国血库协会(AABB)标准保存72小时。经辐照的血小板作为观察组,未辐照的血小板作为对照组。流式细胞术检测两组血小板辐照前及保存24和72小时后P选择蛋白的表达水平;同时用血细胞计数仪检测血小板数和平均血小板体积(MPV)。结果表明:辐照组与对照组血小板表达CD62p百分率均随保存时间的延长而增高;保存24小时与新鲜血小板比较有显著性差异(p(0.05),保存72小时与保存24小时比较有非常显著性差异(p(0.01)。在保存24、72小时后,两组血小板CD62p表达率、血小板计数无显著差异(p0.05);但两组血小板在保存72小时后MPV与新鲜血小板比较有显著性差异(p(0.05)。结论:γ射线照射不影响血小板的数量和质量,但手工血小板液态储存时间应尽可能缩短。     

供者血细胞比容对机采血小板产品计数和收集效率的影响

为提高机采血小板制品的质量，保证临床输血的有效性，人们对血细胞分离机的自动化程度、操作程序以及制品质量等方面不断改进。Fenwal Amicus血细胞分离机是CS-3000plus的换代机型，笔者使用Amicus血细胞分离机采集供者血小板102例，以探讨供者血细胞比容对本机型采集产品的血小板计数、收集效率的影响。     

冰冻机采血小板出现析出物的原因分析与对策

冰冻机采血小板由于贮存期长、使用方便而广泛应用于临床,如由于血小板计数低或功能障碍引起的出血病人的抢救和治疗。冰冻机采血小板因采集制备过程比较复杂,复融过程中出现析出物的情况时有发生。笔者对冰冻机采血小板出现析出物的原因做了观察分析。1材料与方法1.1材料CS3000Plus血细胞分离机及相应耗材(美国Baxter公司);-80℃深低温冰箱(日本SANYO公司);电热恒温水浴箱(苏州医用仪器厂);二甲基亚砜(DMSO,美国),血小板振荡仪(苏州医用仪器厂)。1.2冰冻机采血小板的制备采用CS3000Plus标准血小板单采程序Pro1设定IDO为8,抗凝剂与…     

提高机采血小板血浆氧含量对血小板功能影响的探讨

为了探讨提高机采血小板血浆中的含氧量对血小板功能的影响,将机采血小板样品分为实验和对照2组。实验组在无菌条件下提高机采血小板血浆中含氧量(溶解氧)后,两组同时放置(22±2)℃水平振荡的血小板振荡仪器中保存。分别在血小板保存0、1、2、3、4、5天检测血小板量数量、血小板聚集反应功能、血小板液乳酸含量和血小板CD62p表达量。结果显示2组的血小板数量、血小板聚集反应功能均随保存时间延长而下降;2组间比较,血小板数量无显著性差异(P>0.05),血小板聚集反应功能实验组在保存2-3天明显好于对照组(P<0·05)。2组的血小板乳酸含量、血小板CD62p表达量均随保存时间延长而升高;2组间比较,实验组的血小板乳酸含量和血小板CD62p表达量在保存1-3天时低于对照组(P<0.05);机采血小板的含氧量在0天时实验组显著高于对照组(P<0.01)。结论提高血小板血浆中的含氧量(溶解氧)可弥补保存袋中血小板代谢的供氧不足,提高血小板的有氧代谢和血小板的保存质量。     

两种软件的血细胞分离机采集少白细胞血小板的比较

对两种版本软件血细胞分离机 (COBESpectraLRSSystemVersion 5 .1andVersion 7.0 )采集的少白细胞血小板中白细胞残留量、采集效率、采集速度等进行了比较。使用两种版本软件血细胞分离机共采集 2 32个单位的浓缩血小板 ,其中A组用 5 .1版本软件的血细胞分离机采集 16 3单位血小板 ,B组用 7.0版本加快型软件的机器采集6 9单位血小板。献血者外周血血细胞计数及采集过程中的有关参数、血小板得率、采集效率及血小板制品中残留白细胞数等进行了统计分析。结果表明 ,B组的血小板得率为 (2 .9± 1.1)× 10 11,高于A组的 (2 .5 8± 1.2 )× 10 11。A组 99.4 %血小板制品白细胞污染量小于 5× 10 6,而B组中 97.1%的制品白细胞低于此值。B组的采集效率为(5 1.4± 8.7) % ,高于A组的 (43.6± 6 .3) %。血小板得率与采集前献血者外周血血小板计数相关。结论指出 ,血细胞分离机 7 0加快版本在血小板采集效率上优于 5 .1版本 ,采集前献血者的血小板计数高 ,血小板得率也相应高     

GMA保存血小板的实验研究

目的:探讨葡萄糖甘露醇腺嘌呤(GMA)作为添加剂对血小板的保存效果。方法:将GMA替代血浆制备的浓缩血小板(GMA-PC)和以ACD血浆为介质的浓缩血小板(ACD-PC)置22℃水平摇箱内保存,于各时间段取样测定有关指标。结果:两组PC的血小板低渗休克反应和以ADP诱导的血小板聚集率随保存时间的延长平行降低,差异无显著性(P> 0.05),在保存120h 期间,GMA-PC的乳酸脱氢酶释放量低于ACD- PC,前者的形态积分则显著高于后者,提示用GMA 保存的血小板其形态和细胞膜的完整性优于用ACD血浆保存的血小板。结论:初步证实用GMA替代血浆制备和保存浓缩血小板具有可行性     

以海藻糖为添加剂冷藏保存血小板悬液的研究

本研究探讨以海藻糖(trehalose)为添加剂的血小板悬液冷藏保存方法。采用核素标记法检测血小板生存时间,用比浊法测定血小板聚集率,诱导剂为终浓度11.2μmol/L的ADP。采集兔心脏血,按常规方法制备浓缩血小板悬液(PCs),在悬液中加入50mg/ml的海藻糖,37℃水浴4小时后,放于4-8℃冰箱保存,冷藏12天后,测定血小板聚集功能和输入自身体内后的生存时间。结果表明:常温和冷藏储存24小时后的PC血小板聚集率分别为(75.3±9.8)%和(80.5±12.5)%;输入兔体内24、48和72小时时的血小板存活率分别为(78.1±7.9)%、(65.4±6.7)%、(57.5±7.2)%和(5.1±2.5)%、(2.8±2.0)%、(0.9±0.8)%。加入海藻糖的PC冷藏保存12天后,血小板聚集率为(77.8±9.5)%;输入体内24、48和72小时时的血小板存活率分别为(75.7±11.0)%、(67.0±8.5)%、(56.8±8.0)%,与常温保存24小时对照相比,两者相近,P值均>0.05。结论:海藻糖能保护冷藏储存的兔血小板,延长冷藏血小板在体内的生存时间,经海藻糖冷藏储存的兔血小板功能完好。     

Preparation of leukocyte-poor platelets by filtration

There is evidence that leukocyte contaminating red blood cells and platelet concentrates are responsible for refractoriness to platelet transfusions. The efficacy of a cotton-wool filter to remove leukocytes from red blood cells has been documented previously. The present study was designed to evaluate whether the cotton-wool filters can effectively remove leukocytes from platelet concentrates. Sixty pools of random-donor platelets and single-donor plateletpheresis products were filtered through a cotton-wool filter. The efficacy of filtration was determined by measuring the absolute numbers of leukocytes and platelets and subpopulations of mononuclear cells. The average platelet loss was 8% per pool of random platelets and 10% per plateletpheresis product. The average leukocyte removal was 99% from a pool of random platelets and plateletpheresis concentrates collected by CS-3000 and 90% from plateletpheresis concentrates harvested by single-stage COBE/IBM-2997. The filtration removed 100% of granulocytes, 95% of monocytes, 90% of B-lymphocytes, and 85% of T-lymphocytes. We conclude that filtration through a cotton-wool filter is an efficient and cost-effective method for preparation of leukocyte-poor platelets.     

Platelet collection and transfusion using the fenwal CS-3000 cell separator

A prototype model of the Fenwal CS-3000 Blood Cell Separator (Deerfield, IL) was studied for plateletpheresis in 63 donors and 5 transfusions in patients. Donor effects were consistent with platelet removal and mild hemodilution. The incidence of reactions (9 of 63) was low and all were mild "citrate" type. A two-hour collection yielded 4.0 ± 0.72 × 10(11) platelets at an efficiency of 45 ± 6.9 per cent. The product had little contamination with leukocytes (0.26 ± 1.2 × 10(9) and red blood cells (hematocrit less than 1%). Morphology and pH were well preserved during 24 hours of storage. Four patients with uncomplicated aregenerative thrombocytopenia were transfused on five occasions, with a mean of 4.5 ± 0.87 × 10(11) platelets resulting in a mean platelet count increment of 55,000/microliter and dramatic reduction in template bleeding times.     

Platelet concentrates promote procoagulant activity: evidence from experimental studies using a perfusion technique

BACKGROUND: Evaluation of the hemostatic effectiveness of platelet transfusions is difficult. Perfusion methods have been employed to test the quality and function of platelet concentrates, allowing differentiation between platelet-platelet and platelet-surface interactions. STUDY DESIGN AND METHODS: A study was performed to investigate platelet adhesive and cohesive properties as well as the formation of fibrin when aliquots of platelet concentrates were added to thrombocytopenic blood. Blood previously anticoagulated with low- molecular-weight heparin (20 U/mL) underwent platelet and white cell reduction by filtration. Perfusates were prepared by adding to filtered blood platelets obtained from standard concentrates (stored for 1, 3, and 5 days). The final platelet count in these perfusates was standardized at 80,000 per microL. After perfusions, platelet- subendothelium interaction and fibrin formation were analyzed morphometrically. Results were always compared with those obtained in unfiltered blood (> 150,000 platelets/microL). RESULTS: A slight impairment in the ability of stored platelets to interact with the subendothelium was noticed during the storage period. However, the presence of fibrin was significantly greater than that observed in studies with unfiltered blood (Day 1 = 23.48 +/− 9.43%*; Day 3 = 26.99 +/− 6.74%*; Day 5 = 17.95 +/− 9.06% vs. unfiltered blood = 12.60 +/− 3.08%; *p < 0.05). The lower platelet counts (80,000/microL) in the perfusates containing platelets from concentrates could account for the reduced platelet-subendothelium interactions, but they cannot explain the increments in fibrin formation. CONCLUSION: While the preparation and storage of platelets have a detrimental effect on platelet adhesiveness, such procedures can positively influence the platelet procoagulant activity necessary to platelet hemostasis.     

Platelet concentrates derived from buffy coat and apheresis: biochemical and functional differences

Today, platelet concentrates are generally produced from whole blood by differential centrifugation (buffy coat-derived platelet concentrates--PCs) or by plateletpheresis (apheresis-derived platelet concentrates--APCs). As PCs are characterized by a lower number of platelets than APCs, four to six PCs are customarily combined in order to obtain an equivalent dose. In the 1970s and 1980s, the use of PCs exceeded that of APCs by far; in contrast, since the beginning of the 1990s, APCs comprise more than half of all transfused platelets. However, the selection of PCs or APCs for transfusion to thrombocytopenic patients is still a matter of debate. The present paper compares biochemical and functional properties of both platelet preparations in vitro. Besides plasma parameters (e.g. platelet factor 4 (PF4), P-selectin, C3a-desarginin, plasma coagulation factors), platelet function was analysed by aggregometry and the PFA 100 system. APCs are characterized by a better preservation of ADP and collagen-induced platelet aggregation, and shorter closure times of the PFA 100 test system during storage. The improved primary in vitro haemostatic capacity of APCs is presumed to be owing to a lower cellular activation rate in these preparations. This hypothesis is supported by the higher plasma concentrations of PF4, beta-thromboglobulin and P-selectin found in PCs compared with APCs. The concentrations of C3a-desarginin in PCs exceed those in APCs by far. Additionally, thrombin generation is higher in PCs than in APCs. These data suggest that APCs are characterized by a superior haemostatic capacity over PCs in vitro. However, in vivo studies should be performed to confirm these findings in the patients' circulation also.     

Comparison of plateletpheresis with a standard and an improved collection device

A blood cell separator with a specialized separation chamber ([TNX-6]CS- 3000 Plus) was developed for the collection of platelet concentrates with higher platelet yields and lower white cell contamination than obtained with the standard blood cell separator (CS-3000). To compare these devices, normal donors were scheduled for paired plateletpheresis procedures spaced 4 weeks apart, with one procedure using the CS-3000 Plus and the other using the CS-3000. Overall, the platelet yield per unit (mean +/− SEM) was 4.3 +/− 0.1 × 10(11) with the CS-3000 Plus versus 3.7 +/− 0.1 × 10(11) with the CS-3000 (p < 0.001), and the white cell contamination per unit (mean +/− SEM) with the former was 2.4 +/− 0.7 × 10(6) versus 84.1 +/− 21.1 × 10(6) with the latter (p < 0.001). The sequence of procedures (i.e., the order in which the devices were paired) was selected randomly, and similar results were found regardless of sequence. When donors with predonation platelet counts of > or = 200 × 10(9) per L (n = 21) were studied separately, 76 percent of the collections by the CS-3000 Plus contained > or = 4 × 10(11) platelets versus 34 percent of those by the CS-3000 (p < 0.01), and 93 percent of the collections by the former contained < 5 × 10(6) white cells (69% contained < 1 × 10(6)) versus 0 percent of those by the latter (p < 0.01). Thus, platelet collections with the TNX-6 chamber consistently demonstrated high platelet yields and strikingly low white cell contamination–qualities that justify converting standard devices to devices with a TNX-6 chamber.