ABO血型不合的异基因造血干细胞采集及移植效果研究

本研究评价COBE Spectra血细胞分离机按干细胞采集程序采集HLA配型相合、ABO血型不合供者外周血造血干细胞的效能,观察未去除红细胞和(或)血浆进行异基因外周血干细胞移植的效果。应用COBE Spectra血细胞分离机的自动干细胞采集程序采集28例异基因供者外周血干细胞,并选用同期ABO血型相合15例作对照。检测采集物有核细胞(NC)数、单个核细胞(MNC)比例及CD34+细胞计数,观察造血功能重建情况和转变为供者血型所需要的时间。结果表明,ABO血型不合和相合组采集物中的NC、CD34+细胞数、MNC比例无统计学差异(p>0.05)。ABO血型不合组和相合组中性粒细胞和血小板恢复的时间无统计学差异(p>0.05)。14例ABO血型主要不合患者,红系造血明显延迟,ABO血型不合组28名患者于移植后35-193天血型成功转变为供者型,和ABO血型相合组相比均有统计学差异(p<0.01)。结论:ABO血型不合不是异基因造血干细胞移植的障碍,主要不合可能是红系造血明显延迟的主要原因。     

ABO血型不合供者外周血造血干细胞的采集及移植效果研究

目的探讨应用CS-3000 Plus血细胞分离机采集ABO血型不合供者外周血造血干细胞的效率及不去除红细胞和/或血浆进行异基因外周血造血干细胞移植(PBSCT)的安全性。方法经G-CSF 5μg/(kg.d)动员的异基因外周血干细胞供者33名,应用CS-3000 Plus血细胞分离机的干细胞采集程序于动员后d 5采集,其中ABO血型主要不合12名,次要不合8名以及ABO血型相合13名。根据供者外周血的红细胞压积(Hct)和单个核细胞(MNC)计数,对分离机参数作相应调整。输注前从产品袋中留取干细胞,检测有核细胞数、MNC比例、CD34+细胞数、红细胞、血浆含量。单次处理循环血量(9 986±2 489)ml,抗凝剂用量(971±162)ml。供者采集前注射10%葡萄糖酸钙,以预防低钙反应。观察PBSC输注后受者的生命体征、尿液颜色及是否有溶血相关不良反应等。结果ABO血型主要不合组、次要不合组与ABO血型相合组采集物中的有核细胞数、CD34+细胞数、MNC比例无统计学差异(P>0.05),3组供者每次采集的PBSC产品终体积近60 ml,ABO主要不合组采集物中混入红细胞为(3.67—10.25)×1010/袋,ABO次要不合组采集物中血浆量为22—38 ml,不去除红细胞及血浆,直接回输给受者,均未出现溶血反应,所有患者造血功能均获得重建。结论应用CS-3000 Plus血细胞分离机采集ABO血型不合供者的外周血干细胞,通过调整分离机参数,减少ABO血型不合红细胞的混入,可以获得足够的干细胞数量并安全用于移植。     

Peripheral blood stem cell collection in allogeneic donors: impact of venous access

BACKGROUND: Peripheral blood stem cell (PBSC) collection is accepted as a routine procedure in related and unrelated healthy donors worldwide. Venous access can be accomplished by peripheral veins or a central venous catheter (CVC). STUDY DESING AND METHODS: We compared efficacy and tolerability of 40 PBSC collections via CVC with 6267 PBSC collections via peripheral veins in healthy allogeneic donors. Results of the leukapheresis procedures and side effects in the donors were evaluated. RESULTS: The median CD34+ cell counts on Day 5 and the results of the stem cell collection were not significantly different between the two groups of allogeneic donors. Pain or problems at the site of puncture or catheter insertion occurred in 58.6% of the donors with a CVC versus 37.8% of the donors with peripheral venous access (p = 0.03). The incidence and severity of paresthesia during the leukapheresis was not significantly different in both groups of donors (p = 0.09). During follow‐up no major adverse events related to CVC were reported. CONCLUSION: Central femoral lines proved to be safe and tolerable in healthy allogeneic donors but peripheral venous access should be preferred, whenever possible.     

Peripheral blood mononuclear cell collection from patients undergoing adoptive immunotherapy or peripheral blood-derived stem cell transplantation and from healthy donors.

The demand for collection of mononuclear cells from the peripheral blood of patients for therapeutic purposes is rapidly increasing. Automated blood cell separators are usually designed for collection of blood components from healthy donors. We reviewed safety and efficiency of collection data of a new procedure for the Fenwal CS 3000 blood cell separator in 125 collections from normal donors and 101 collections from patients after IL-2 pretreatment or chemotherapy. The new procedure set red blood cell spillovers to occur at 3.5 minute intervals, using procedure 1 with the interface detector set at 1,000 and the standard granulocyte and collection chambers. Despite significant anemia and thrombocytopenia in a large number of patients no serious procedure-related side effects occurred. The lymphocyte yield was 4.74 +/- 1.6 x 10(9) per 5 liters of blood processed in normal donors and 24.2 +/- 12.0 x 10(9) per 10 liters of blood processed after IL-2 treatment. After chemotherapy the lymphocyte yield was 4.5 +/- 3.1 x 10(9) per 10 liters of blood processed; the collection efficiency was found to be significantly lower in this group. The main problem was the platelet loss of 35.6 +/- 12% of the initial count in normal donors, 40.3 +/- 14.1% after IL-2 treatment, and 42.1 +/- 18.0% after chemotherapy. The platelet loss is, however, closely related to the preapheresis platelet count; patients with thrombocytopenia lose fewer platelets than normal donors. Therefore the procedure was found to be safe for patients with a platelet count as low as 20/nl. This report provides a basis for safe, effective mononuclear cell collection from patients with very abnormal peripheral blood counts.     

Peripheral blood stem cell collection for allogeneic hematopoietic stem cell transplantation: Practical implications after 200 consequent transplants

Background

Proper stem cell mobilization is one of the most important steps in hematopoietic stem cell transplantation (HSCT). The aim of this paper is to share our 6 years’ experience and provide practical clinical approaches particularly for stem cell mobilization and collection within the series of more than 200 successive allogeneic HSCT at our transplant center.

Analysis of factors associated with successful allogeneic peripheral blood stem cell collection in healthy donors

BackgroundThe collection of a sufficient number of stem cells is important for success of allogeneic hematopoietic stem cell transplantation (HSCT). This study aimed to investigate the factors associated with successful allogeneic peripheral stem cell (PBSC) collection in healthy donors.MethodsWe retrospectively reviewed clinical data of allogeneic PBSC collection in 175 donors from 2007 to 2017 at the National Cancer Center, Korea. This study analyzed factors associated with the CD34+ cell yield such as the characteristics of donors, including age, laboratory results before apheresis, and data of procedures on the first day. The CD34+ cell dose of ≥ 4.0 × 10⁶/kg have recently been the accepted minimum recommended dose in allogeneic HSCT settings, and this was the target dose in our study.ResultsThe factors associated with the CD34+ cell yield were age (p = 0.007), baseline platelet (PLT) (p = 0.014), and pre-collection hematopoietic progenitor cells (HPCs) (p = 0.001) by multivariate analysis. This study represented that age, baseline platelet count, and pre-collection HPC count are important predictive factors as shown in other previous studies.ConclusionOur data suggest that young age, high baseline platelet counts and high HPC counts before collection might be useful for identifying successful mobilizers.     

G‐CSF mobilized peripheral blood stem cell collection for allogeneic transplantation in healthy donors: Analysis of factors affecting yield

Mobilized PBSC are the main source for allogeneic HSCT. We aimed to evaluate factors that affect CD34+ cell yield including the donor's age, gender, BSA, processed blood volume and the method of G‐CSF dose calculation. Data from 170 healthy donors were analyzed. The concentration of CD34+ cells in the peripheral blood (PB) and the processed volume of blood were significantly correlated to CD34+ cells yield (P < .00005 and P < .001, respectively). The G‐CSF dose per m² was significantly correlated to the concentration of CD34+ cells in the PB (P = .0003) and in the product (P = .01). Smaller BSA and less processed volume were found among female donors, who were given lesser G‐CSF dose per m², and showed lower yield compared to men. However, multivariate analysis of the yield showed that only the concentration of CD34+ cells in the PB and the processed volume remained independent significant.     

健康供者外周血造血干细胞动员后采集的最佳时机

背景:外周血干细胞移植的首要条件是造血干细胞有效动员和采集,并能较快持久地重建造血.近年国内外不少单位已开展了此项工作,但平均每例供者采集次数较多,且造血干细胞的产率高低差别较大. 目的:探讨健康供者外周血造血干细胞动员后采集的最佳时机.方法:2003-01/2008-12海口市人民医院进行异基因外周血造血干细胞采集的健康捐赠者16例,随机分为2组,单纯动员组6例,皮下注射粒细胞集落刺激因子5.0～10.0 μg/(kg·d),共5 d;联合组10例,在单纯动员组基础上,静脉推注地塞米松10 mg.每例供者均采集2次外周血造血干细胞,单纯动员组与联合组又各分为2 h,4 h两个亚组,即若在第4天动员后2 h采集,则第5天在动员后4 h采集;若在第4天动员后4 h采集,则第5天在动员后2 h采集.16例供者采集量为4.0～5.0 mL,推赶量3.0～5.0 mL,总循环血量6.7～10.1 L.结果与结论:单纯动员组、联合组采集的单个核细胞数均达到(4.0～8.0)×10~8 kg~(-1).与2 h亚组比较,两组4 h亚组采集物的单个核细胞数均明显增加(P < 0.05).提示粒细胞集落刺激因子动员后第4,5天进行采集,4 h采集物的细胞浓度明显高于2 h;供者采集时循环血量是其自身循环血量的1.8～2.2倍时已能一次采集足够量的细胞数,有较合理的采集时间-价值-效益的关系.     

异基因造血干细胞移植健康供者外周血干细胞二次采集时机探讨

目的 探讨健康供者体内应用rhG-CSF初次采集造血于细胞以后二次外周血造血干细胞采集的时机.方法 38例二次采集外周血干细胞的健康供者皮下注射rhG-CSF 5 μg·kg-1·d-1,连用5 d,在第5、6天采集外周血移植物(A组);A组供者初次采集时的资料作为对照(B组);A组供者依据初次和二次采集的75%间隔时间分为C组(≤9个月,n=30)和D组(>9个月,n=8).应用流式细胞术检测各组供者外周血采集物中的淋巴细胞,CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-T细胞的数量.结果 A组供者外周血采集物中淋巴细胞CD3+CD8+(25.51×108)和CD34+细胞(0.51×108)的中位含量显著低于B组(31.55×108和0.70×108,P<0.05);C组供者CD3+CD8+(23.42×108)和CD34+细胞(0.42×108)的中位含量显著低于B组(P<0.05);D组供者淋巴细胞,CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-T细胞的中位含量与B组的差异无统计学意义(P>0.05).A、C和D三组采集物中CD4+与CD8+细胞的比值、单核细胞与CD3+细胞的比值以及CD3+CD4-CD8-T细胞与CD3+细胞的比值与B组的差异均无统计学意义(P>0.05).38名健康供者初次和二次采集的间隔时间与二次采集物中的CD34+细胞存在显著的相关性(r=0.357,P=0.028).结论 健康供者二次采集外周血造血干细胞的时机以初次采集的9个月后为宜,9个月内采集应适当增加循环血量以保证移植物中的免疫和造血组分能满足临床需要.     

AutoPBSC程序与MNC程序采集外周血造血干细胞的比较研究

目的比较COBE Spectra血细胞分离系统的自动采集程序(AutoPBSC程序)与4.7版半自动采集程序(MNC程序)采集健康供者外周血造血干细胞的差异及对供者血常规相关指标的影响。方法 2002年3月~2008年3月期间对53例健康供者随机采用Auto PBSC程序和MNC程序进行了113例次造血干细胞采集,其中采用AutoPBSC程序63例次,MNC程序50例次。分析比较2种程序采得外周血造血干细胞(PBSC)的采集体积、单个核细胞百分数及总数、CD34+细胞百分数及总数等指标,采集前后供者红细胞、血小板变化。结果 2种程序采集外周血造血干细胞的体积、单个核细胞百分数、CD34+细胞百分数、CD34+细胞总数、采集袋中血小板及红细胞混入量、采集前后供者血小板计数的变化存在显著性差异(P<0.01);2种程序采集的单个核细胞总数无显著性差异;血小板计数在应用MNC程序组呈下降趋势,较AutoPBSC程序更加明显(P<0.01);1例地中海贫血供者应用Auto PBSC程序采集失败。结论 2种程序均可有效采集外周血造血干细胞,与MNC程序比较AutoPBSC程序具有以下优势:单个核细胞和CD34+细胞百分数提高、采集物体积减少有利于采集物的冻存、采集物中血小板数少对供者血小板影响小。地中海贫血供者需慎用Auto PBSC程序。     

非亲缘异基因造血干细胞移植供者的护理

造血干细胞移植(hematopoietic stem cell transplantation)为近40年新兴的临床应用科学,已广泛应用于急性白血病、慢性白血病、淋巴瘤等多种疾病的治疗,而异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation)具有复发率低,采集的干细胞浓度高等优势,因此在临床已被广泛应用[1].     

非亲缘异基因造血干细胞移植供者的护理

造血干细胞移植(hematopoietic stem cell transplantation)为近40年新兴的临床应用科学,已广泛应用于急性白血病、慢性白血病、淋巴瘤等多种疾病的治疗,而异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation)具有复发率低,采集的干细胞浓度高等优势,因此在临床已被广泛应用[1].     

儿童外周血造血干细胞采集

目的　探索儿童外周血干细胞 (PBSC)的采集 ,特别是体重 <2 0kg的小儿外周血干细胞采集方法。方法　 18例儿童 ,共 31次采集PBSC。①年龄小 ,血管细和不能很好地配合的儿童患者 ,在PBSC采集的前 1d ,根据患者的情况 ,选择性行锁骨下静脉、股静脉或颈内静脉穿刺 ,插入一儿童型双腔管 (18～ 36cm) ;②采集儿童PBSC时 ,使用SVSC分离槽和SVCC收集槽 ,体重 <2 0kg的小儿 ,在初始化完成后 ,使用经射线照射过的血液预先运转分离机 ,使儿童体内的血液保持平衡 ;③在分离过程中 ,ACD/全血的比例保持在 1∶11～ 1∶13之间 ;④成人的PBSC采集一般循环量为 10 0 0 0～ 15 0 0 0ml,儿童为全身血容量的 2～ 3倍 ,每次处理血液总量为 4 80 0～ 10 0 0 0ml。结果　除 2例采集 3次外 ,其余的经 1～ 2次PBSC采集 ,均收集到足够的MNC和CD3 4 细胞 ,达到外周血干细胞移植的阈值要求。结论　使用本方法能很好地采集儿童外周血造血干细胞。     

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)     

Peripheral blood stem cell collection and transplantation using the Haemonetics Multi Component System

AIM: To assess clinical usefulness of an intermittent-flow blood cell separator in peripheral blood stem cell (PBSC) collection and transplantation. RESULTS: The Haemonetics Multi Component System (Multi) was used to collect PBSC (52 aphereses in 17 patients). The mean processing blood volume and the mean PBSC yield were 7407 ml and 2.16 x 10(6) CD34+ cells/kg, respectively. When CD34+ cells exceeded 0.3% of the peripheral WBC, more than 2.0 x 10(6) CD34+ cells/kg could be collected by a single apheresis. Eight patients underwent PBSC transplantation after high-dose chemotherapy. Hematopoietic recovery was achieved in a median period of 10 days. CONCLUSIONS: (1) A single-arm, light-weight machine has sufficient capability to collect PBSC. (2) The percentage of CD34+ cells in the peripheral WBC is a good predictor of the CD34+ cell yield of the collection.     

Peripheral blood hematopoietic stem cell mobilization and collection efficacy is not an independent prognostic factor for autologous stem cell transplantation

BACKGROUND: The successful mobilization and collection of hematopoietic stem cells are dependent on a number of clinical factors such as previous chemotherapy and disease stage. The aim of this retrospective study was to determine whether the effectiveness of mobilization and collection is an independent prognostic factor for autologous stem cell transplantation outcome. STUDY DESIGN AND METHODS: A total of 358 patients who received transplants from January 2003 to December 2004 (201 male and 157 female patients, ages from 2.7 to 77.3 years with median of 53 years of age) underwent autologous hematopoietic stem cell collection after mobilization with granulocyte-colony-stimulating factor (G-CSF) or G-CSF plus chemotherapy priming. This retrospective study included patients with diagnoses of acute myelogenous leukemia, non-Hodgkin's lymphoma, Hodgkin's disease, multiple myeloma, and solid tumors. All patients underwent stem cell collection until a target or a minimum CD34+ cell dose was reached. Correlations were performed between stem cell mobilization and/or collection efficacy and transplantation outcomes. RESULTS: In general, both larger reinfused CD34+ cell dose and shorter number of days for the stem cell count to reach the minimum of 2 x 10(6) per kg CD34+ cells do not foster quicker engraftment. Reinfused CD34+ cell dose of less than 12 x 10(6) and number of days stem cell collection to reach this minimum CD34+ cell dose did not independently affect the overall survival (OS) or disease-free survival (DFS). CONCLUSION: The effectiveness of hematopoietic stem cell mobilization and collection as defined as number of days to reach a CD34+ cell dose of 2 x 10(6) per kg should not be used independently to forecast posttransplantation prognosis, engraftment, DFS, and OS.     

Collection of two peripheral blood stem cell concentrates from healthy donors

When peripheral blood stem cell (PBSC) concentrates are used for allogeneic transplants, two or more apheresis procedures must often be performed. To determine how many cells could be collected from healthy people by two back-to-back apheresis procedures and what effect these collections would have on donors, we gave 19 healthy people 5 micrograms kg-1 day-1 and 21 people 10 micrograms kg-1 day-1 of granulocyte colony stimulating factor, filgrastim, for 5 days. We then collected two PBSC concentrates, one on day 5 and one on day 6. A third group of six people was given filgrastim 10 micrograms kg-1 day-1 for 5 days but had no PBSC concentrates collected. PBSC concentrate cell counts and donor cell counts, symptoms, and blood chemistries were assessed for up to 1 year. On day 5, three times more CD34+ cells were collected from donors given 10 micrograms kg-1 day-1 than those given 5 micrograms kg-1 day-1 (P = 0.009) but on day 6 the quantity of cells collected was the same (P = 0.23). The total number of CD34+ cells collected was two times greater in donors given the higher dose of filgrastim (median = 579 x 10(6); range = 174-1639 x 10(6) compared to 237 x 10(6); 103-1670 x 10(6); P = 0.061). Platelet counts fell after each PBSC concentrate collection, but there were no differences between the two groups of donors in platelet counts measured immediately after each collection. The platelet counts also fell in people who did not donate PBSC concentrates. The lowest counts in all three groups of people also occurred on day 10. In PBSC donors given 10 micrograms kg-1 day-1 of filgrastim the absolute neutrophil count (ANC) fell below premobilization counts on day 14. In donors given 5 micrograms kg-1 day-1 the ANC fell below premobilization counts on days 21, 28 and 49, CD34+ cell counts were significantly lower than premobilization counts on days 14 and 28 in donors given 10 micrograms kg-1 day-1 of filgrastim and on day 14 in those given 5 micrograms kg-1 day-1. No decrease in neutrophil or CD34+ cell counts occurred after filgrastim was given in the people who did not donate PBSC concentrates. The incidence of symptoms was similar in both groups of PBSC concentrate donors, except that those given 10 micrograms kg-1 day-1 were more than twice as likely to experience myalgias as those receiving the lower dose (P = 0.029). Several blood chemistries changed. Levels of alkaline phosphatase, LDH, SGPT, SGOT, uric acid and sodium increased. Levels of bilirubin, total protein, potassium, calcium and chloride decreased. In conclusion, twice as many CD34+ cells were collected from donors given 10 micrograms kg-1 day-1 of filgrastim. Platelet, neutrophil and CD34+ cell counts fell after the PBSC concentrate collections. The fall in platelet counts was due to both the collection and the administration of filgrastim. The falls in neutrophil and CD34+ cell counts were due to the loss of haematopoietic progenitor cells in the PBSC concentrates. Allogeneic PBSC concentrate donors should be given 10 micrograms kg-1 day-1 of filgrastim, and if possible only one component should be collected in order to avoid thrombocytopenia.