Peripheral blood stem cell collection with a blood cell separator

Forty-three patients with malignant nonmyeloid diseases underwent peripheral blood stem cell collections on an apheresis system (Spectra, COBE BCT, Lakewood, CO). Collections took place during the white cell (WBC) recovery phase following conditioning chemotherapy. One hundred two procedures were done after chemotherapy alone, and 72 procedures after chemotherapy plus granulocyte-colony-stimulating factor (G-CSF). Four centrifugal separation factors were tested. One and one-half patient blood volumes were processed in each procedure. The mean volume of the collected component was 158 +/− 16 mL. After chemotherapy alone, the procedures provided a mean of 0.8 × 10(8) WBCs per kg and 2.3 × 10(4) colony-forming units-granulocyte macrophage (CFU-GM) per kg of recipient body weight. The mononuclear cell percentage in the components increased with the centrifugal separation factor from 85 to 96 percent. In parallel, platelet contamination increased from 2.1 to 3.8 × 10(11). The collect hematocrit ranged from 1.0 to 2.5 percent (0.01-0.025). The collection efficiency for mononuclear cells and CFU- GM also increased with the centrifugal separation factors from 52 to 70 percent for mononuclear cells and from 55 to 68 percent for CFU-GM. Collections performed after G-CSF-stimulated mobilization were characterized by a higher neutrophil contamination independent of centrifugal separation factor, which gave a mean mononuclear cell percentage of 64 percent in the collected component. The average yield for these procedures was 2 × 10(8) WBCs per kg and 28 × 10(4) CFU-GM per kg.(ABSTRACT TRUNCATED AT 250 WORDS)     

全自动血细胞分析仪与血细胞形态学检验在血常规检验中的应用价值

目的比较全自动血细胞分析仪和血细胞形态学检验在血常规检验中的应用价值。方法选择2019年6月至2019年7月在本院进行血常规检验的3 000例患者为研究对象,将所有患者的样本均分为A、B两份,分别采用全自动血细胞分析仪和全自动血细胞形态学数字图像分析仪检查。以人工显微镜检查结果作为金标准,比较两种检验方式的白细胞异常检测结果。结果 A组检测结果显示,3 000例患者样本的阳性率为3.67%,灵敏度为90.99%,特异度为99.69%;B组检测结果显示,3 000例患者样本的阳性率为3.67%,灵敏度为97.30%,特异度为99.93%。B组检测的灵敏度高于A组(P<0.05)。结论全自动血细胞形态学数字图像分析仪在白细胞异常检测中的灵敏度高于全自动血细胞分析仪,具有效率高、速度快的优势,但仍不能替代人工镜检。     

Red blood cell storage and cell morphology

Aim: In this study, we performed weekly assessment of morphology‐related parameters through monitoring of CPD‐SAGM leuco‐filtered erythrocyte concentrates from blood withdrawal until the 42nd day of storage. Background: Liquid storage of red blood cells (RBCs) delivers a blood‐derived therapeutic, which is safe, available, effective and affordable for most patients who need transfusion therapy in developed countries. However, a growing body of accumulating controversial evidences, from either biochemical or retrospective clinical studies, prompted safety concerns about longer stored RBCs. Methods: Statistical image analysis through scanning electron microscope was coupled to osmotic fragility and erythrocyte sedimentation rate. Results: We could observe that by day 21 more than 50% of RBCs displayed non‐discocyte phenotypes. This observation was related to an increase in osmotic fragility, which was totally overlapped in day 0 controls and day 7 RBCs while only slightly augmented in day 14 samples. Cation dysregulation (pH internal/external alteration and potassium) might both reflect and trigger a negative feedback loop with metabolic fluxes and membrane cation pumps. Conclusion: Morphology parameters suggest that significant alterations to RBC morphology over storage duration occur soon after the 14th day of storage, as to become significant enough within the 21st day.     

Red blood cell alloimmunization among sickle cell Kuwaiti Arab patients who received red blood cell transfusion

BACKGROUND: Sickle cell disease (SCD) is common in the Arabian Gulf region. Most cases require a red blood cell (RBC) transfusion, increasing the potential for RBC alloantibody development. The incidence of RBC alloimmunization among Kuwaiti Arab SCD patients is not yet known. This study retrospectively assessed the effect of using two different matching protocols on the incidence of alloimmunization among multiply transfused Kuwaiti Arab SCD patients.
STUDY DESIGN AND METHODS: A total of 233 Kuwaiti Arab SCD patients were divided into two groups: Group 1 (n = 110) received RBC transfusion through standard ABO- and D-matched nonleukoreduced blood; Group 2 (n = 123) received RBCs matched for ABO, Rh, and K1 poststorage-leukoreduced blood. Multivariate analysis was performed on the factors associated with RBC alloimmunization and antibody specificity.
RESULTS: Sixty-five percent of patients in Group 1 developed clinically significant RBC alloantibody with an increased prevalence in females; in patients in Group 2, 23.6% developed RBC alloantibodies (p = 0.01). In Group 1, 72 patients (65.5%) had alloantibodies directed against Rh and Kell systems (p = 0.01). Multivariate analysis further confirmed the results, showing that blood transfusion type and sex have significant effects on the rate of alloimmunizations.
CONCLUSION: This study confirms the importance of selecting RBCs matched for Rh and Kell to reduce the risk of alloimmunizations among Kuwaiti Arab SCD patients.     

血细胞分析仪自动计数外周血有核红细胞的应用研究

目的评价血细胞分析仪自动计数外周血有核红细胞(NRBC)的方法学特点,并探讨其临床应用价值。方法选择115例血液病及非血液病患者,用SysmexXE2100全自动血细胞分析仪及显微镜计数法计数其外周血中NRBC数量。结果SysmexXE2100与显微镜计数法计数外周血中NRBC数量有极好的相关性(r>0.97),并且2种方法的计数结果差异无显著意义(P=0.2018)。高值、中值和低值NRBC标本绝对计数的批内CV平均<6.5%。在(0～15)×109/L范围内计数NRBC有极好的线性(r=0.9998)。SysmexXE2100自动计数外周血中NRBC的灵敏度和阴性预测值均为100%。外周血中NRBC增高时,WBC显著高于去除NRBC的计数结果(P=0.0168)。结论SysmexXE2100血细胞分析仪可灵敏、准确、精密地自动计数外周血中NRBC,并且可在NRBC增高的标本中准确计数白细胞数量。除白血病、贫血等血液病患者外,非血液病(如肿瘤化疗)等患者外周血中也可出现NRBC。血细胞分析仪自动计数外周血中NRBC,有助于一些血液病及非血液病患者的筛查与诊断。     

Comparison of two blood cell separators in collecting peripheral blood stem cell components

To ensure that a sufficient number of CD34+ cells are collected for an allogeneic blood progenitor cell transplant, the most effective blood cell separator should be used to collect peripheral blood stem cell (PBSC) components. We compared the effectiveness of two blood cell separators. We gave 29 healthy people 7.5 or 10 μg kg^?1 of granulocyte colony stimulating factor (G-CSF) daily for 5 days and collected one PBSC component with either a Fenwal CS3000 (n = 15) or a Cobe Spectra (n = 14) blood cell separator. The volume of blood processed was the same for each machine (8.4 ± 1.0 L; range = 4.9–9.4 L for the CS3000 and 8.9 ± 1.0 L; range 6.7–10.9 L; P = 0.71). The components collected with the CS3000 contained more mononuclear cells (39.6 ± 21.9 × 10⁹ compared with 26.9 ± 5.6 × 10⁹, P = 0.02) and fewer neutrophils (1.38 ± 1.88 × 10⁹ compared with 5.53 ± 8.71 × 10⁹, = 0.001). The total number of CD34+ cells collected with the two instruments was the same (470 ± 353 × 10⁶ for the CS3000 and 419 ± 351 × 10⁶ for the Spectra; P = 0.64) as was the number of CD34+ cells collected per litre of whole blood processed (55.9 ± 42.0 × 10⁶ L^?1 compared with 45.9 ± 37.9 × 10⁶ L^?1; P = 0.59). The mononuclear cell collection efficiency was greater for the CS3000 (82.4 ± 54.9% compared with 53.3 ± 14.1; P = 0.04) but the CD34+ cell collection efficiencies were the same (87.4 ± 61.1% for the CS3000 compared with 56.3 ± 23.5% for the Spectra; P = 0.07). In conclusion, both blood cell separators collected components which contained large numbers of CD34+ cells, but those collected with the CS3000 contained fewer neutrophils and the CS3000 was more efficient at collecting mononuclear cells.     

Preparation of packed red blood cell units in the blood bank: Alteration in red blood cell deformability

BackgroundDonated blood is stored in the blood bank as packed red blood cell units. In the process of packed cells preparation, the red blood cells (RBCs) are subjectedto high level of shear stress, which can induce alterations in their properties.In the present study, we examined the effect of packed RBCs preparation (which included leuko-filtration) on red cell deformability.MethodsBlood samples were collected from 25 healthy donors and from corresponding units of packed RBCs. The portion of undeformable cells (%UDFC) was determined for each sample.ResultsThe median value of %UDFC was equal to 6.75 ± 0.70 %, for freshly-donated RBCs, and to 6.36 ± 0.51 %, for packed cells. Wherein, %UDFC may increase or decrease following packed cells preparation, depending upon the initial portion of undeformable cells.ConclusionLikely, exposure of RBCs to high shear stress, during packed cells preparation, induces opposing effects: (a) removal/destruction of rigid (undeformable) cells, thereby reducing their total amount (i.e., decreasing the %UDFC) on the one hand, and (b) mechanical damage to the cell membrane and subsequent reduction of the cell deformability (thereby increasing the %UDFC) on the other. As a consequence, the final impact of packed cells preparation is primarily determined by the initial state of erythrocytes in the blood of the donor.     

致敏红细胞和未致敏红细胞物理特性比较

目的研究致敏红细胞和未致敏红细胞形态大小及渗透脆性的差异。方法对健康献血者0型和Rh（＋）型红细胞用IgG型抗-D致敏后,与未致敏的相同献血者的红细胞用血细胞分析仪进行检测,比较两种红细胞的平均红细胞体积和红细胞分布宽度,用红细胞渗透脆性试验对二者进行渗透脆性比较。结果致敏红细胞平均体积大于未致敏红细胞（P〈0.05）,未致敏红细胞分布宽度大于致敏红细胞（P〈0.01）,致敏红细胞渗透脆性大于未致敏红细胞。结论红细胞致敏后在体积上大于未致敏红细胞,渗透脆性降低,可能与红细胞膜上血型抗原与相应抗体结合后对红细胞膜的结构和稳定性有一定的影响。     

Red blood cell transfusion therapy

This article describes the characteristics of alloantibodies associated with accelerated in vivo red cell destruction, pretransfusion tests performed to detect these antibodies, and the resolution of the incompatible crossmatch. Current transfusion practices, the indications for, and the complications of red cell transfusions are discussed.     

SYSMEX XN-3000血液分析仪检测外周血有核红细胞的应用评估

目的评价SYSMEX XN-3000血液分析仪(简称XN-3000)计数外周血有核红细胞(NRBC)的方法学特性和临床应用价值。方法分别用XN-3000与人工显微镜镜检法对96例血液标本进行检测,采用SPSS 11.0统计软件分析其相关性。结果 XN-3000检测NRBC的敏感性为98.41%,特异性为93.94%,假阳性率为3.13%,假阴性率为3.13%,检测结果重复性好,低值变异系数(CV)为6.5%,中、高值CV均5%。与显微镜镜检法计数外周血中NRBC数量有极好的相关性(r20.867),且两者差异无统计学意义(P=0.019)。结论使用XN-3000进行外周血NRBC计数,方法可靠,操作简便快速,准确性高,可替代显微镜法,有一定的临床应用价值。     

两种血细胞分离机分离外周血造血干细胞效果比较

目的　比较FenwalCS 3000plus及CobeSpectra两种血细胞分离机分离外周血造血干细胞的效果。方 法　共有76名供者入组。FenwalCS 3000plus组31例,平均年龄(39.1±12.9)岁,共循环38次;CobeSpectra组 45例,平均年龄(38.2±11.2)岁,共循环65次。比较两组采集物白细胞总数、单核细胞百分率、采集物单核细胞中 CD34+细胞百分率。结果　两组采集物的白细胞计数无明显统计学差异;FenwalCS 3000plus组单核细胞百分率 高于CobeSpectra组。两组采集物中CD34+细胞百分率无明显统计学差异。CobeSpectra组采集物中单核细胞百 分率与白细胞数负相关。两组单核细胞百分率、两组白细胞数均与CD34细胞百分率无相关性。结论　本组资料 显示在分离外周血干细胞方面FenwalCS 3000plus与CobeSpectra之间没有差异。     

Red blood cell endothelial nitric oxide synthase does not modulate red blood cell storage hemolysis

BACKGROUND: The red blood cell (RBC) endothelial nitric oxide synthase (eNOS) has been shown to regulate intrinsic RBC rheologic properties, such as membrane deformability, suggesting that a functional eNOS could be important in RBC viability and function during storage. This study examines the correlation between RBC eNOS deficiency and the propensity of RBCs to hemolyze under selected stress conditions including prolonged hypothermic storage. STUDY DESIGN AND METHODS: Fresh or stored RBCs from normal and eNOS knockout (KO) mice or from healthy human volunteers were subjected to selected hemolytic stress conditions including mechanical stress hemolysis, osmotic stress hemolysis, and oxidation stress hemolysis and evaluated during standard storage in CPDA‐1 solutions. RESULTS: Fresh RBCs from normal and eNOS KO mice demonstrated comparable susceptibility to hemolysis triggered by mechanical stress (mechanical fragility index 6.5 ± 0.5 in eNOS KO vs. 6.4 ± 0.4 for controls; n = 8‐9), osmotic stress, and oxidative stress. Additionally, RBCs from both mouse groups exhibited similar hemolytic profile at the end of 14‐day hypothermic storage, analogous to 42 days of human RBC storage. Storage of human RBCs (28 days in CPDA‐1) in the presence of NOS cofactors (l ‐arginine and tetrahydro‐l ‐biopterin) or inhibitor (N⁵‐[imino(methylamino)methyl]‐l ‐ornithine monoacetate) did not affect cell recovery or hemolytic response to the selected stressors. CONCLUSION: These studies suggest that RBC eNOS does not modulate susceptibility to hemolysis in response to selected stress conditions or prolonged hypothermic storage. Other strategies to increase nitric oxide (NO) bioactivity after prolonged storage utilizing NOS‐independent pathways such as the nitrate–nitrite–NO pathway may prove a more promising approach.