不同方法制备的血小板在保存期内活化状态研究

目的研究白膜回浆法（Bc）制备的浓缩血小板和血细胞分离机（Trima）制备的单采血小板在保存期内的活化状态。方法采用流式细胞术和酶联免疫吸附法分别检测BC和Trima制备的血小板在5d保存期内血小板表面CD62P和血浆可溶性cD62P（scD62P）的表达。结果在O～5d的保存期内,两种方法制备的血小板表面CD62P和血浆sCD62P的表达均随保存时间推移而增加;BC制备的浓缩血小板CD62P阳性率和血浆sCD62P的表达均明显高于Trima制备的单采血小板。结论BC制备的浓缩血小板活化程度高于Trima制备的单采血小板,其制备工艺需改进,以减少血小板活化水平。     

不同型号血细胞分离机对献血者细胞参数与机采血小板采集效率比较

目的比较不同型号血细胞分离机对献血者细胞参数与机采血小板采集效率。方法选取2018年1月至2019年11月我站的318例献血者,其中159名献血者应用AmiCORE、MCS+及Trima血细胞分离机捐献双份血小板,平均分为3组,另外159名献血者应用AmiCORE、MCS+及Trima血细胞分离机捐献单份血小板,平均分为3组。比较三组的单份及双份血小板细胞参数不合格率及分离机性能。结果MCS+组单份血小板细胞参数不合格率低于AmiCORE组和Trima组,差异具有统计学意义(P<0.05)。MCS+组单份血小板采集效率及时间高于AmiCORE组及Trima组,差异具有统计学意义(P<0.05);Trima组抗凝剂使用量及全血处理量多于MCS+组及AmiCORE组(P<0.05)。三组双份血小板细胞参数不合格率比较,差异均无统计学意义(P>0.05)。MCS+组双份血小板采集效率高于AmiCORE组及Trima组,差异具有统计学意义(P<0.05);AmiCORE组采集时间长于MCS+及Trima组,Trima组抗凝剂使用量及全血处理量多于MCS+及AmiCORE组,差异具有统计学意义(P<0.05)。结论AmiCORE、MCS+及Trima血细胞分离机对产品采集具有较高采集效率,符合国家有关标准,血液制备和采集过程可供,各机型血小板细胞参数不合格率、血小板分离机性能存在区别。血液制备及血液采集过程具有可控性,应依据献血者自身特点选择不同血细胞分离机。     

冰冻保存单采血小板聚集原因探讨

目的：探讨-80℃冰冻保存单采血小板聚集原因，避免聚集的发生。方法：用CS3000-PLUS血细胞分离机采集血小板。按冰冻单采血小板操作规程制备冰冻血小板。回顾分析冰冻血小板的聚集情况。结果：制备冰冻血小板518袋。解冻后发生肉跟可见的血小板聚集10袋，其中使用进口产品袋和转移袋冰冻保存血小板聚集发生率分别为0．47％和0．35％；使用国产产品袋和分浆袋冰冻保存血小板聚集发生率分别为55．6％和27．27％。血小板聚集与冰冻保存时间关系不大。讨论：使用进口产品袋和进口转移袋冰冻保存血小板发生聚集率(0．47％和0．35％；)均低于使用国产产品袋和分浆袋(55．6％和27．27％)；提示使用进口袋冰冻保存血小板可明显降低冰冻血小板聚集率。国产袋出现血小板聚集多的原因可能是：(1)由于血袋含有某些杂质，加入冰冻保护剂DMSO后溶解进入血小板悬液，引起血小板聚集；(2)血袋透气性较差。内壁不够光滑，使血小板的代谢消耗增加和机械磨擦刺激，而导致血小板损伤；(3)由于透气性较差，使速冻过程延长，而导致血小板冰冻机械损伤。     

单采血小板献血不良反应原因分析及对策

目的 通过分析单采血小板献血者相关资料,探讨单采血小板献血不良反应原因,采取有效对策减少献血不良反应的发生.方法以美国血液技术公司生产的MCS3P、MCS+血细胞分离机采集的902例为Ⅰ组,Trima血细胞分离机采集的1 315例为Ⅰ组.结果两组间献血不良反应发生原因的差异有统计学意义.结论根据献血者的红细胞比容、血管...     

血小板校准系数对单采血小板质量的影响

目的 通过比较单采设备中血小板校准系数(YSF系数)调整前、后单采血小板含量,探讨YSF系数对单采血小板质量的影响.方法 将480例无偿献血者分成2组,每组240例.选取Trima Accel血细胞分离机,采集240例无偿献血者当前YSF系数的单采血小板.按照操作手册调整Trima Accel系统的YSF系数,采集调整YSF系数后240例无偿献血者的单采血小板.比较YSF系数调整前、后两组的血小板收集量、血小板容量、白细胞计数、红细胞计数及储存期末pH值等的差异.结果 YSF系数调整前、后两组血小板收集量、白细胞计数比较,差异有统计学意义(P<0.05);但血小板容量、红细胞计数及储存期末pH值比较,差异无统计学意义(P>0.05).调整后单采血小板的血小板含量不合格率由24.67％降至13.33％.结论 调整YSF系数可降低单采血小板含量不合格率,提高单采血小板的质量,采供血机构应制订标准工作流程,规范YSF系数的设置,以保证临床输注的疗效.     

低温-80℃长期保存单采血小板的功能研究

目的 :为明确长期冰冻保存的单采血小板的功能 ,并对冰冻单采血小板的保存时间进行初步探讨。方法 :对冰冻保存前后的单采血小板进行回收率、粘附率、聚集强度和Ⅲ因子活性进行检测。结果 :冰冻保存后的回收率为 83 8%± 8 3 % ,粘附率为 5 9 0 %± 9 2 % ,聚集强度为 66 1%± 13 2 % ,Ⅲ因子活性为 ( 18± 3 )s。结论 :冰冻保存后的单采血小板仍具有较好的功能活性     

献血者血小板数量及血容量对机采血小板采集量的影响

目的：分析献血员血小板数量及处理血量对机采血小板采集量的影响。方法应用Trima血细胞分离机采集100名献血者的双份血小板。结果通过多元回归分析发现,献血者采前的血小板计数（Plt）和血容量（BV）与血小板采集量之间存在回归关系,其标准偏回归系数β值分别为-0.370和-0.201,P＜0.01。结论对采前的Plt和BV水平较低的献血者,可采用Trima血细胞分离机进行采集。     

三种血细胞分离机单采血小板洗涤效果及临床应用比较

目的随机选取三种型号血细胞分离机（CS-3000plus、Mcs、Trima）采集的浓缩血小板，作为制备洗涤血小板的原料，对血小板回收率、血浆蛋白清除率、临床应用效果进行比较。方法分别取浓缩血小板（PCs）和富血小板血浆（PRP）为原料，PCs直接加入0．9％Nacl 200ml洗涤、离心、去上清；再加入0．9％ Nacl悬浮；PRP要先离心去除血浆，再按上述同样方法洗涤。检测每袋洗涤血小板回收率和血浆蛋白清除率，并对临床应用效果进行比较。结果CS-3000 Plus血细胞分离机采集血小板的终产品是浓缩血小板，洗涤后血小板平均回收率为95．16％；而Mcs和Trima血细胞分离机采集血小板的终产品是富血小板血浆，洗涤后血小板平均回收率分别为89．06％和89．84％；三者血浆蛋白清除率均大于98％；洗涤血小板的的质量达到国家《全血及成分血质量标准》中单采血小板和洗涤血细胞的质量标准，临床疗效明显，并可预防输血过敏反应。结论洗涤血小板采用浓缩血小板作为洗涤原料，比富血小板血浆回收率高，操作更为简便。     

冰冻血小板制备研究

[目的]探讨冰冻单采血小板制备的可行性,为其临床应用提供可靠依据.[方法]通过认真选择献血员,单采血小板,并对40袋新鲜血小板进行计数等质控,用二甲基亚砜（DMSO）作冰冻保护剂,-80℃深低温保存1年内,解冻后观察外观,分别进行计数、计算回收率、测定PH值、粘附率、无菌试验,并且与新鲜血小板进行比较.[结果]-80℃保存冰冻单采血小板1年内血小板计数、pH值、粘附率与冻前新鲜血小板进行比较差异无显著性（P〉0.05）;血小板平均体积、血小板体积分布宽度冰冻前后差异有显著性（P〈0.01）,保存1年内平均回收率为90.49% , 无菌实验无细菌生长.[结论]-80℃深低温保存冰冻血小板质量达到标准要求,可以应用于临床.     

血小板保存期间血小板计数和P-选择蛋白的变化

探讨CS 30 0 0plus血细胞分离机全密闭 5天保存袋保存的血小板数量和质量的变化 ,研究在 (2 2± 2 )℃条件下振荡和静置两种方法保存的血小板有无差异。采用SysmexF 82 0型全自动血细胞分析仪对机采血小板产品计数 ,应用酶联免疫吸附法 (ELISA)定量检测机采血小板上清液中P 选择蛋白的含量。结果显示 ,振荡与静置两种方法保存的血小板产品中血小板计数和P 选择蛋白含量均无显著性差异。随保存时间延长 ,振荡和静置保存的血小板计数逐渐减少和P 选择蛋白逐渐增多 ,其中两组内的血小板计数在保存 0 - 72小时以内均无显著性差异 ;而在保存 96 - 1 2 0小时后与保存前比较都有显著性差异。振荡保存组 0 - 72小时内 ,每相邻 1天之间P 选择蛋白量无显著性差异 ,其余各时间段均有显著性差异 ;静置保存组 0 - 72小时内各时间段均无显著性差异。结论 :①使用CS 30 0 0plus血细胞分离机全密闭 5天保存袋 ,对血小板制品的有效保存期以 3天为宜 ;②在 (2 2± 2 )℃条件下 ,振荡保存的血小板在数量和质量上并不优于静置保存的血小板。     

全血手工制备浓缩血小板后的血浆再制备冷沉淀的质量评价

目的评价室温新鲜全血白膜法制备浓缩血小板后的血浆再制备冷沉淀的质量。方法实验组为24例,新鲜全血(400 mL)置室温于<8 h用白膜法制备浓缩血小板后所得的血浆,冰冻保存。对照组1为12例,常规制备新鲜冰冻血浆,冰冻保存。对照组2为12例,新鲜冰冻单采血浆,血浆单采完毕分装为200 mL/袋并立即冰冻保存。3组血浆按常规制备冷沉淀,评价其质量:外观、凝血因子FⅧ及Fib的含量;血细胞残留量。结果 3组冷沉淀外观均正常;WBC含量在3组间无统计学意义。与对照组1比较:实验组凝血因子FⅧ(81.76±34.07)IU较低,Fib(202.63±48.58)mg及Plt(7.81±5.81)×109均较高。与对照组2比较:实验组凝血因子FⅧ含量相当,Fib(202.63±48.58)mg较高、Plt(7.81±5.81)×109较低。结论全血来源的制备浓缩血小板后的冰冻血浆还可以用于冷沉淀的制备,其质量符合国家标准。     

A comparison of frozen and fresh platelet concentrates in the support of thrombocytopenic patients

Ten patients scheduled to receive intensive chemotherapy were plateletapheresed and the platelet-rich plasma was frozen with 5 percent dimethyl sulfoxide at -80 to -95 degrees C until needed. Paired comparisons of frozen autologous platelets with fresh single-donor platelets were made in seven patients using corrected platelet increments at 1 and 24 hours, and pre- and posttransfusion bleeding times. In vitro tests of 12 units of platelet-rich plasma before and after freezing included platelet factor 4 (PF4) secretion, malondialdehyde production, and electron microscopic evaluation of morphology. Fresh platelets provided significantly better 1- and 24-hour corrected increments compared with frozen autologous platelets. In only one case of alloimmunization did frozen autologous platelets provide a better increment than fresh platelets. Bleeding times after transfusion showed no consistent improvement regardless of type of transfusion or platelet count. Secretable PF4 remained constant after freezing, but malondialdehyde production fell significantly. Platelets showed considerable structural damage with 33 percent balloon forms counted after thawing, compared to less than 1 percent before freezing. Except in the case of alloimmunization, frozen autologous platelets are inferior to single-donor fresh platelets, and are significantly damaged in the freezing process.     

Comparative Studies of the Viability of Human Platelets Stored at 4 C in Plastic and Glass Containers

Studies were done to investigate whether the surface of the container (plastic or glass) would influence the viability of human platelets stored at 4C for short intervals.
Cr⁵¹-labeled platelets prepared as concentrates suspended in plasma were preserved at 4 C for 24, 48 or 72 hours and their capacity to recirculate and survive after infusion into the respective donors ("viability") was determined. In addition, the clot retraction property of the stored platelets was measured.
Platelet viability was sharply reduced after storage at 4 C in all experiments. When the storage period was limited to 24 hours, survival curves of platelets preserved in plastic bags were similar to those of platelets preserved in glass bottles. However, after 48 hours of storage, viability of platelets preserved in plastic containers had values significantly higher than those of platelets preserved in glass containers. After 72 hours of storage, platelet viability was reduced to minimal values but was still greater for platelets stored in plastic bags.
The study of clot retraction confirmed data previously obtained and showed that this platelet property was preserved better by storage in glass bottles rather than in plastic bags. Platelet viability was, however, lost very rapidly during storage at 4 C in either type of container so that the unfavorable effect of plastic on the preservation of clot retraction was thought not to be of practical importance in platelet transfusion therapy.     

An in vivo comparison of CPD and CPDA-2 preserved platelet concentrates after an 8-hour preprocess hold of whole blood

To see if citrate-phosphate-dextrose-adenine-two (CPDA-2) anticoagulant- preservative had an effect on the viability of platelets, we studied autologous in vivo recovery and survival in humans for platelet concentrates prepared from six units of blood drawn into CPDA-2 and compared them to six units drawn into citrate-phosphate-dextrose (CPD). These units were prepared from whole blood held at room temperature for 8 hours after collection and were then stored for 3 days at 22 ± 2 degrees C. The recovery for platelets preserved in CPD was 39.0 +/0 4.8 percent and for platelets preserved in CPDA-2, 32.5 ± 4.4 percent. The difference was not significant (p greater than 0.10). In order to estimate population differences, in vitro effects on in vivo viability were also evaluated. Six in vitro variables were studied but only pH at 72 hours (r = 0.77), platelet count (r = 0.64), and morphology score (r = 0.66) correlated to recovery. Only pH at 72 hours significantly influenced recovery (p = 0.007). By adjusting for individual pH differences, mean recovery for platelets stored in CPD was 37.5 percent, and for platelets stored in CPDA-2, 34.0 percent. The mean lifespan was 6.7 ± 0.7 days for platelets preserved in CPD and 6.1 ± 1.0 days for those preserved in CPDA-2. Although hemostatic function was not studied, these data support in vitro observations that platelets preserved with CPDA-2 are not different from platelets preserved with CPD, even after 8-hours of storage of whole blood at room temperature prior to platelet concentrate preparation.     

改良白膜法制备浓缩血小板及回收率的影响因素

背景：白膜法和富含血浆法制备的浓缩血小板有无效输注发生率高和不良反应发生率高的缺点。 目的：观察改良白膜法制备浓缩血小板的实验研究,分析制备浓缩血小板回收率的影响因素。 方法：随机抽取126例站内采集后4-6 h的400 mL血液,随机分成改良白膜法组、白膜法组和富含血浆法组。改良白膜法采用3步离心,第1次采用次重离心,离心转速2300 r/min,离心时间12 min,降速5,离心温度（22±2）℃;第2次采用轻离心,离心转速910 r/min,离心时间10 min,离心温度（22±2）℃;第3次离心转速2800 r/min,离心时间12 min,离心温度（22±2）℃,离心后,挤去上层含血小板较少的血浆,袋中留30 mL血浆悬浮血小板,即为浓缩血小板。通过数据库文献检索的方法分析制备浓缩血小板回收率的影响因素。 结果与结论：改良白膜法、白膜法以及富含血浆法制备的手工浓缩血小板中,制备前各组血小板总数差别无统计学意义（P 〉0.05）;富含血浆法组和改良白膜法组较白膜法组血小板回收率高,差异有显著性意义（P 0.05）;白膜法组和改良白膜法组较富含血浆法组残留红细胞和残留白细胞的量少,差异有显著性意义（P0.05）。制备浓缩血小板的回收率受到全血量、离心转速、离心时间、离心方法等因素的影响。改良白膜法制备浓缩血小板减少红细胞和白细胞的残留量,提高了血小板的回收率,可在血液中心或中心血站推广应用。     

Evaluation of the performance of Trima Accel® v5.2 for the collection of concentrated high-dose platelet products and concurrent plasma from high platelet count donors, in Germany

Background and objectives: This study was undertaken to test the ability of Trima Accel® version 5.2 to simultaneously collect concentrated high‐dose leukoreduced platelet products and double doses of plasma. Materials and methods: Random volunteers (18–65 years of age) with preprocedure platelet counts above 270 × 10³/μl were recruited among the blood center's apheresis donors. All complied with the center's donor selection criteria. Results: One hundred fourteen (114) collections were performed. Depending on which definition of single platelet dose is used (2.0 × 10¹¹ as prevalent standard in most European countries, and 3.0 × 10¹¹ as prevalent standard in the United States and Canada) in 107/114 (single dose = 2.0 × 10¹¹) and 39/114 (single dose = 3.0 × 10¹¹) instances, a triple platelet product was obtained. In 87 cases (76%), a double plasmaproduct (>430 ml) was collected, and in seven cases (6%), a single plasma product (>220 ml) was collected. In 20 procedures, only platelets without concurrent plasma were collected (18%). Overall procedure time was 87 ± 13 min and average platelet yield per procedure was 8.5 ± 1.4 × 10¹¹ (final storage concentration, 1,279 ± 153 × 10³/μl). The median residual leukocyte content per transfusion dose was 0.13 × 10⁶ (0.02–0.98 × 10⁶) for a single dose of 2.0 × 10¹¹ and 0.14 × 10⁶ (0.02–0.98 × 10⁶) for a single dose of 3.0 × 10¹¹. Conclusions: Trima Accel® version 5.2 allows for collection of concentrated high yield platelet products. It offers high productivity and reliably achieves the configured yield targets. Leukoreduction performance complied with both US and EU legal requirements. Collection as hyperconcentrates furthermore allowed for concurrent collection of double dose plasma in the majority of the procedures. J. Clin. Apheresis, 2012. © 2012 Wiley Periodicals, Inc.     

Platelets and factor VIII as functions of blood collection time

Plasma and platelet concentrates were prepared from whole blood collected in less than eight minutes, and from blood collected in 8 to 12 minutes. Comparison was made between the two collection groups of Factor VIII activity in fresh plasma, and of Factor VIII activity in fresh frozen plasma. Using the rank sum test, no significant difference was found between Factor VIII activities in either fresh or fresh frozen plasma. The number of platelets in concentrates prepared from blood collected in less than eight minutes were compared with those in concentrates from blood collected in 8 to 12 minutes. Mean numbers of platelets were 6.89 × 10(10) and 7.12 × 10(10),respectively. No significant difference was found between the two groups when compared by the rank sum test, or the "t" test. Current guidelines recommending maximum eight-minute collection for component preparation may be inappropriately restrictive.     

冰冻血小板融化复苏后不可逆聚集的原因分析

目的探讨冰冻血小板融化复苏后不可逆聚集的原因.方法对血小板收集浓度、二甲基亚砜融化温度、采后震荡与否、加入速度以及融化过程进行分组观察. 结果血小板收集浓度不同,不可逆聚集发生率的差异有显著性(P＜0.01);DMSO融化温度、加入速度对血小板质量有显著影响(P＜0.01);血小板采后震荡可明显降低冻融后不可逆凝集率(P＜0.01).结论冰冻血小板采集、制备、冻融等各个环节必须严格质量控制,规范操作.