脐血造血干／祖细胞研究进展

脐血和骨髓均富含造血干／祖细胞，但二者在造血干／祖细胞的含量上存在差别。脐血中各CD34^ 细胞亚群有着独特的免疫表型。脐血造血干／祖细胞能在体外扩增，扩增后的CD34^ 细胞能在SCID小鼠体内造血，由脐血CD34^ 细胞可以生成淋巴细胞和内皮细胞。脐血造血干／祖细胞移植与骨髓造血干／祖细胞移植在患者的生存率、复发率及移植免疫排斥等方面均具有可比性，有着极大的潜在价值。     

血细胞分析仪快速检测外周血造血干细胞与祖细胞的临床意义

目的 探讨血细胞分析仪快速检测外周血造血干细胞/祖细胞(hematopoietic stem/progenitor cell,HPC)的临床应用价值.方法 方法学验证和回顾性分析.收集2015年1月至2018年12月期间来福建医科大学附属协和医院就诊的4例为初治急性髓系白血病患者外周血、1名健康供者外周血干细胞采集物5...     

凋亡在脐血造血干/祖细胞冷冻损伤中的作用及其机制研究

为探讨脐血造血干／祖细胞冷冻损伤中凋亡的作用及其机制，应用流式细胞术检测脐血CD34^ 细胞冻存前后细胞凋亡率、线粒体膜势能和Fas抗原，用免疫组织化学法检测Bcl—2蛋白表达，用Western blot和caspase-3蛋白活性分析检测caspase-3的表达．以甲基纤维素半固体培养进行造血祖细胞集落分析。研究结果显示，CD34^ 细胞在-196℃冻存2周或4周后凋亡率增加，电泳出现DNA梯形条带，CFUs分别下降了25．2％和30．1％。冷冻过程中细胞内线粒体膜势能下降，Bcl—2蛋白表达下调，而Fas抗原表达无明显改变。新鲜分离的CD34^ 细胞中caspase-3以分子量为32kD的非活性酶原形式存在，冷冻过程中caspase-3被激活，可检测到分子量为20kD的裂解片段，caspase-3蛋白活性分别增加了1．2倍和1.5倍。结论：凋亡在脐血造血干细胞冷冻损伤中起着重要作用，其机制可能是通过线粒体介导的caspase依赖性途径执行，caspase-3是其中一个重要的效应蛋白酶。     

两种血细胞分离机干/祖细胞采集和移植效果的比较

背景：外周血造血干/祖细胞通过血细胞分离机进行体外收集,血细胞分离机的性能、工作状况将直接关系到采集物的特性和数量,从而直接影响受者的造血重建。目的：比较Fenwal CS-3000Plus与Amicus两种血细胞分离机采集外周血造血干/祖细胞及受者移植效果。方法：共入选接受异基因外周血干细胞移植患者51例,Fenwal CS-3000Plus组27例,平均年龄（34.2±10.6）岁;Amicus组24例,平均年龄（35.4±12.1）岁。比较两组采集物有核细胞数、CD34＋细胞数、采集效率、红细胞与血小板采集量及造血重建时间。结果与结论：两组采集物的有核细胞数、CD34＋细胞数、采集效率、红细胞采集量及造血重建时间差异均无显著性意义;Fenwal CS-3000Plus组血小板采集量明显高于Amicus组（P〈0.01）。结果显示Fenwal CS-3000Plus与Amicus血细胞分离机在分离外周血干/祖细胞方面和移植效果没有差异;血小板较低被采集者使用Amicus血细胞分离机采集可能更为安全。     

外周血CD34+细胞计数预测造血干细胞收获量

目的了解外周血CD34+细胞计数与造血干细胞收获量的关系.方法用流式细胞仪Pro-COUNT方法检测了16例造血干细胞移植患者单采前外周血和采集物中CD34+细胞百分率、绝对数,用血细胞计数仪计数白细胞数.分析各项检测指标与CD34+细胞收获量(×106/kg体重)的相关性.结果外周血白细胞计数、CD34+百分率、CD34+细胞计数与CD34+细胞收获量的相关系数分别为-0.307,0.738,0.665.经相关系数t检验,单采前外周血白细胞计数与CD34+收获量无相关性(P＞0.05),而外周血CD34+百分率和绝对计数与CD34+收获量密切相关(P＜0.01).结论外周血CD34+计数可准确预测采集效果,确定干细胞采集时机.     

Cobe Spectra与Fenwal CS 3000 Plus血细胞分离器外周血造血干/祖细胞的采集效能比较

为了评价Cobe Spectra(Version6.1)和Fenwal CS 3000 Plus两种血细胞分离机在造血干／祖细胞采集中的采集效能，对36人64次外周血造血干／祖细胞的采集进行了回顾性分析．20人42次使用Cobe Spectra采集，16人22次使用Fenwal CS 3000 Plus采集对CD34^ 细胞采集量、采集效率以及红细胞与血小板的采集量进行比较，结果表明：Cobe Spectra和Fenwal CS 3000 Plus在CD34^ 细胞采集量和采集效率上无显著差异?CD34^ 细胞采集量与供者白细胞、单核细胞、造血祖细胞、CD34^ 细胞的动员情况呈正相关，在多因素stepwise线性回归模型中，采集前外周血干／祖细胞浓度是唯一有意义的影响因素，Fenwal CS 3000 Plus的采集效率与外周血单核细胞量呈负相关。Fenwal CS 3000 Plus对红细胞的收集量显著高于Cobe Spectra，并且采集后供者外周血血小板降低程度较Cobe Spectra更严重。结论：Cobe Spectra(Version 6．1)和Fenwal CS 3000 Plus在CD34^ 细胞的采集能力上无显著差别。当外周血单核细胞数量高，或是血型不合移植的及血小板减少的供者，推荐使用Cobe Spectra血细胞分离机.     

脐血造血干/祖细胞的定量检测

脐血造血干细胞移植具有来源广泛、采集便利和移植后移植物抗宿主反应低等优点 ,目前国内已有单位在筹建脐血造血干细胞库。脐血中造血干细胞定量检测对于造血干细胞移植有重要的指导意义 ,由于各地的检测方法和实验条件不同 ,导致所得结果差异较大。本研究应用流式细胞仪 ( FCM) ,探讨 3种检测方法的优缺点 ,并对收集的 61 5份脐血进行造血干 /祖细胞定量检测 ,现报告如下。材料与方法1　仪器与试剂　 ACT diff血细胞计数仪 ( Coulter公司 ) ,FACSCalibur流式细胞仪 ( BD公司 )。CD34/CD38方案中 CD45抗体为 BD产品 ,其余为 Phar…     

两种血细胞分离机干/祖细胞采集和移植效果的比较

背景:外周血造血干/祖细胞通过血细胞分离机进行体外收集,血细胞分离机的性能、工作状况将直接关系到采集物的特性和数量,从而直接影响受者的造血重建.目的:比较Fenwal CS-3000 Plus与Amicus两种血细胞分离机采集外周血造血干/祖细胞及受者移植效果.方法:共入选接受异基因外周血干细胞移植患者51例,Fenwal CS-3000 Plus组27例,平均年龄(34.2±10.6)岁;Amicus组24例,平均年龄(35.4±12.1)岁.比较两组采集物有核细胞数、CD34+细胞数、采集效率、红细胞与血小板采集量及造血重建时间.结果与结论:两组采集物的有核细胞数、CD34+细胞数、采集效率、红细胞采集量及造血重建时间差异均无显著性意义;Fenwal CS-3000Plus组血小板采集量明显高于Amicus组(P<0.01).结果显示Fenwal CS-3000 Plus与Amicus血细胞分离机在分离外周血干/祖细胞方面和移植效果没有差异;血小板较低被采集者使用Amicus血细胞分离机采集可能更为安全.     

CD34^＋造血祖细胞定向诱导分化为T／NK细胞的研究进展

成熟T／NK细胞具有重要的移植免疫和抗肿瘤免疫效应。利用造血干／祖细胞具有多向分化的潜能，通过体外培养体系（含促进T／NK细胞分化发育的以下成份：如SCF、Flt-3L、IL－2、IL-7、IL-12、IL-15、TNF-α等细胞因子，胸腺肽／胸腺素，胸腺基质细胞等不同组合），定向诱导其分化为成熟的T／NK细胞，可用于造血干细胞移植后的细胞过继免疫治疗。本文就CD34^ 造血祖细胞定向诱导分化为T／NK细胞的实验研究的理论基础和进展作一综述。     

SE-9000型血液分析仪在检测外周血干细胞中的临床应用

目的探讨SE-9000型血液分析仪在外周血造血干细胞检测的临床应用价值.方法采用SE-9000型血液分析仪与流式细胞仪(FCM)对1例脐带血干细胞和2例骨髓干细胞移植患者的造血干细胞连续6 d进行检测.结果两种检测方法所得数据差异无显著性,具有良好的相关性(r=0.923,P<0.01).结论 SE-9000型血液分析仪为临床提供了一种快速简捷、费用经济的造血干细胞检测方法.     

SE—9000型血液分析仪在检测外周血干细胞中的临床应用

目的　探讨SE 90 0 0型血液分析仪在外周血造血干细胞检测的临床应用价值。方法　采用SE 90 0 0型血液分析仪与流式细胞仪 (FCM)对 1例脐带血干细胞和 2例骨髓干细胞移植患者的造血干细胞连续 6d进行检测。结果　两种检测方法所得数据差异无显著性 ,具有良好的相关性 (r =0 .92 3,P <0 .0 1)。结论　SE 90 0 0型血液分析仪为临床提供了一种快速简捷、费用经济的造血干细胞检测方法     

Selective enrichment of hematopoietic progenitor cells from peripheral blood

We tried to improve progenitor cell yields by different modifications on conventional leukocyte separation protocols to obtain a more selective stem cell enrichment from peripheral blood. Our data suggest that especially higher process volumes allow selective increases of progenitor cell recoveries. Differences in stem cell yields according to different malignancies are discussed.     

XE-2100血液分析仪与流式细胞仪在造血干/祖细胞检测中的相关性研究

目的探讨利用XE-2100全自动血液分析仪(简称XE-2100)检测脐血中造血干细胞(HPC)数量的可行性。方法利用XE-2100的幼稚通道(IM I)和流式细胞仪(FCM)分别对50例脐静脉血进行HPC计数。结果在脐血中,XE-2100检测HPC数量与FCM检测的CD34 细胞数量有良好的相关性(r=0.89),批内变异系数(CV)<5%。结论XE-2100能快速简便地动态监测脐血HPC的变化,从而为HPC数目的测定提供一种新的测定方法,为临床快速判断HPC数目提供依据。     

Collection of peripheral blood progenitor cells with an automated leukapheresis system

BACKGROUND: Apheresis devices designed for the collection of mature blood elements are being used for the collection of peripheral blood progenitor cells (PBPCs).The collection of PBPCs differs from that of other cells in the rarity of the target cell and in the fact that donors may undergo several days of collection. A consequence of this process may be a depletion of blood cells such as platelets from the blood. The disposable set and operating software for an apheresis device (Spectra, COBE BCT) was modified by the manufacturer to automate the collection of PBPCs and reduce the collection of unwanted blood cells. STUDY DESIGN AND METHODS: A study was initiated to compare the collection of PBPCs with the new device, the AutoPBSC (version [V]6.0 with AutoPBSC tubing set), and that with the MNC (mononuclear cell) procedure (V4.7 with white cell tubing set), for patients and healthy donors. RESULTS: Patients whose blood was processed by either theV6.0 orV4.7 procedure achieved the target dose of 5 x 10(6) CD34+ cells per kg of patient weight in similar numbers of procedures, even though the calculated collection efficiency for CD34+ cells using the automated V6.0 procedure was significantly less than that with the V4.7 procedure for both allogeneic donors and patients donating PBPCs. The collection efficiency for platelets was lower with the V6.0 procedure, and components collected in this manner contained fewer platelets. Apheresis by the V6.0 procedure required 30 to 60 more minutes per procedure than apheresis by the V4.7 procedure. Review of engraftment kinetics after transplantation did not reveal any effect of the collection procedure on recipients of either allogeneic or autologous transplants. CONCLUSION: The collection efficiencies of the V6.0 procedure for both CD34+ cells and mature blood cells are lower than those of the V4.7 procedure.The lower collection efficiency for platelets results in a smaller drop in peripheral blood platelet count after the procedure.The automated features of the V6.0 procedure may simplify PBPC collection, but this procedure requires a longer apheresis.     

Automatic analysis of normal bone marrow blood cells using the XE-2100 automated hematology analyzer

The bone marrow aspiration test conventionally has been performed by visual methods, using a light microscope, because automatic blood cell analyzers cannot adequately capture erythroblasts and immature granulocytes (IGs) (Tatsumi et al.: Osaka City Med J 1988;34:135-146; Tatsumi et al.: Am J Clin Pathol 1986;86:50-54). With the development of the XE-2100 automatic blood cell analyzer (Sysmex Corporation, Kobe, Japan) in 1999, the classification of erythroblasts and IGs by means of flow cytometry (Zini et al.: Infus Ther Transfus Med 2001;28:277-279; Briggs et al.: Sysmex J Int 1999;9:113-119) became possible. In the present study we classified cells in 65 bone marrow aspiration specimens by the microscopic method and with the XE2100, and compared the results. A good correlation was found in the nucleated red blood cell (NRBC), white blood cell (WBC), and total nucleated cell (TNC) counts; the myeloid/erythroid ratio (M/E ratio); neutrophils, lymphocytes, eosinophils, and IGs in the immature myeloid information (IMI) channel; and the total cell count. These items can all be analyzed in about 54 sec with the XE2100, which is faster than with the microscopic method. Therefore, analysis of bone marrow aspiration specimens with this apparatus appears to be very useful for clinical screening as well as laboratory testing.     

Association of cord blood platelet characteristics and hematopoietic progenitor cells

BACKGROUND: Total nucleated cell (TNC) dose is associated with neutrophil and platelet (PLT) engraftment after cord blood (CB) transplantation and thus is used for selection of CB for banking. The goal of this study was to evaluate the internal relationships of CB PLT characteristics, TNC, and the hematopoietic progenitor cell (HPC) content of CB units. STUDY DESIGN AND METHODS: HPC and TNC counts of 167 CB units processed with an automated cell separation system were compared with CB PLT count and mean PLT volume (MPV). Megakaryocyte progenitors (CFU-MK) were cultured from a subset of units (n = 24). RESULTS: PLT concentration correlated with MPV (r = -0.39), which was also associated with both TNC and total CD34+ cells before and after processing (r = 0.37 and 0.35 and r = 0.41 and 0.42, respectively). In addition, MPV was associated with HPC counts in the CB unit. The p value was less than 0.001 for all associations. PLT count was inversely associated with markers of hematopoietic potential. Median removal of PLTs during processing was 62 percent (range, 40%-84%). All 24 CB units of the subset exhibited CFU-MK growth. In multivariate linear regression analysis, MPV improved prediction of the HPC content of the CB unit compared to prediction with CB volume and nucleated cell concentration only. CONCLUSION: Mean PLT volume correlated with current markers of CB hematopoietic potential and is potentially useful for evaluating CB collections for banking. The question of the clinical significance of PLT characteristics in CB transplantation remains unanswered.     

BC-5500全自动血细胞分析仪性能评估报告

目的对新购进的迈瑞BC-5500全自动血细胞分析仪的各项性能进行系统评估并汇总报告。方法按照国际血液学标准化委员会（ICSH）制定的评价标准进行。结果各参数携带污染率皆符合ICSH小于1％的要求且线性良好（r≥0．995）;总重复性除血红蛋白（HB）,平均红细胞血红蛋白量（MCH）略微超出外,其他各项均满足1／4美国临床检验修正法规（CLIA’88）要求;在与华西医科大学附属第二医院检验科所使用XE-2100及本科室所使用XE-2100比对中,除平均红细胞血红蛋白浓度（MCHC）的相关性欠佳外,各参数均有良好的相关性（r≥0．975）;批间精密度各参数均满足1／3 CLIA’88要求;除手动低、中值MCH和MCHC略微超出外,批内精密度均满足1／4C LIA’88要求;各项参数在与华西医科大学附属第二医院检验科所使用XE-2100及本科室所使用XE-2100比对中均在偏倚允许范围内;白细胞分类的重复性及准确性均符合要求。结论本机各项评估试验均达到评估要求,可以接受使用。对于个别结果欠佳的参数,将在日常使用及下次评估中继续观察。     

不同自动血液分析仪血细胞参考区间比较

引言血细胞计数和白细胞分类计数广泛应用于临床实践,适当的参考区间值对理解病人的检测结果是十分必要的。在以前的研究中用来建立相关参考值的分析仪被采用新的技术和能提供额外参考值的仪器所取代。新近介绍的血液分析仪包括能够提供网织红细胞绝对数量和其成熟指数的模式。最近进行的研究是使用新型五种自动血液分析仪对一组西欧成年人进行测试以确定相关的参考区间。主题,材料和方法主题和样本我们的研究议定书遵从NCCLS和1FCC的建议,研究方案完全符合医学伦理制度法案.所有的志愿者都是知情同意。志愿者通过在参与医院、当地大学…