Bone marrow and peripheral blood stem cell harvesting.

A number of malignant diseases are responsive to supralethal myelotoxic high-dose chemoradiotherapy, and can be treated with hematopoietic stem cell rescue. The number of genetic diseases correctable by replacement of defective pluripotent stem cells with normal stem cells or through gene transfer is ever-increasing. In all these cases, pluripotent hematopoietic stem cells need to be infused, which can be obtained directly from the bone marrow or from the peripheral blood with or without the aid of mobilizing strategies. This article is a detailed review of the technical and medical aspects of stem cell collection from the bone marrow and the peripheral blood.     

外周血造血干细胞采集期间低钙血症的预防与治疗

背景：在造血干细胞采集过程中需以复方枸橼酸钠抗凝全血，枸橼酸盐可鳌合血液中的钙离子，形成不能离子化的枸橼酸钙而引起血液中钙离子下降，引起低钙血症。目的：探寻在外周血干细胞采集期间预防及治疗低钙血症的最佳方法。设计、时间及地点：析因设计的观察对比实验，于1998—10／2007-12在华中科技大学附属协和医院血液科完成。对象：共纳入168例外周血造血干细胞采集患者，其中健康异基因外周血造血干细胞供者122例，自体外周血造血干细胞移植46例。方法：采集前皮下注射粒细胞集落刺激因子进行干细胞动员，至第5天应用Cobe Spectra血细胞分离机的自动外周血干细胞采集程序进行外周血造血干细胞采集，平均循环血量为8410mL，分离过程中平均采血速度为42mL／min。163例供者采集2次，5例采集3次，共341次。根据补钙方案分为2组，口服组102例供者（共209例次采集）口服葡萄糖酸钙；静脉组66例（共132例次采集）静脉持续滴注葡萄糖酸钙。主要观察指标：采集期间观察两组供者低钙血症相关症状的发生，采集完毕后从产品袋中留取标本计算有核细胞数和CD34^＋细胞计数。结果：①口服组209例次的供者采集中，43例次（20．6％）出现如口周、四肢麻木感、胃肠道反应、胸闷、头昏等的低钙血症表现，经加大口服葡萄糖酸钙剂量或静脉注射100g几葡萄糖酸钙症状均得到控制；静脉组132例次的供者采集中，5例次（3．8％）出现低钙血症表现，经加大静脉注射100g／L葡萄糖酸钙症状亦得到控制。口服组供者低钙血症的发生率显著高于静脉组（P〈0．01）。②口服组和静脉组采集物中的有核细胞数、CD34^＋细胞计数差异无显著性意义（P〉0．05）。结论：应用血细胞分离机采集外周血造血干细胞时，采用静脉持续滴注葡萄糖酸钙补钙的方法明显优于口服葡萄糖酸钙，且不影响造血干细胞的采集效率。     

An alternative method for custom prime: A case report of successful peripheral blood stem cell harvesting from two low-weight child donors

Allogeneic peripheral blood stem cell (APBSCs) transplantation is an effective treatment for hematological malignancies. However low-weight donor children meet some complications. In the current report, PBSCs were harvested from a 14-month-old child (9.8 Kg) for a 6 years old sibling recipient suffering from pre-B type of acute lymphoblastic leukemia (ALL) and also 24 months old male child donor (12 Kg) for a haploidentical recipient suffering from acute myeloid leukemia (AML-M4EO). The PBSC harvesting was performed using Spectra? Optia^® apheresis software with continuous mononuclear cell (CMNC) procedure. The results were completely promising and both recipients underwent an acceptable transplantation.     

Predictive factors for successful peripheral blood stem cell mobilization and collection in children

Factors affecting the success of peripheral blood stem cell collection (SCC) in children are not well characterized. We reviewed 218 stem cell collections among 199 pediatric donors, of which 35 were from healthy sibling donors and 164 were for autologous collections. Successful SCC, defined as a CD34⁺ cell count of ≥2 × 10⁶/kg of recipient weight per intended transplant, occurred in 188 of 199 donors (94%). Ideal SCC defined ≥5 × 10⁶ CD34⁺ cells/kg of recipient per intended transplant, occurred in 147 (74%) patients. Failure of collection occurred in 11 (6%) patients and was significantly associated with an autologous collection for a brain tumor diagnosis (P = .003) and a pre-apheresis peripheral blood (PB) CD34⁺ count <20 × 10⁶ cells/L (P = .002). Ideal SCC was significantly associated with age < 10 years (P = .01) and pre-apheresis PB-CD34⁺ count ≥20 × 10⁶ cells/L (P < .0001). Factors associated with failure of SCC may be identified in advance of the collection procedure allowing appropriate counselling of patients as well as anticipatory guidance for multiple collections or justify the preemptive use of stem cell mobilizing agents.     

Evaluation of the Fresenius cell separator AS 104 for harvesting peripheral blood stem cells in pediatric patients

In a single institution trial we carried out 35 peripheral blood stem cell harvesting procedures in 12 children with advanced malignancies to evaluate the procedure's safety and the collection efficiency of the Fresenius blood cell separator AS 104 in a pediatric population. Despite a significant mean decrease of 21% (+/-8%) in systolic blood pressure after starting the procedure, all children tolerated leukapheresis without any adverse reaction. After termination of leukapheresis there was a significant decrease of all determined hematological parameters, as compared with pre-harvest values. The mean mononuclear cell recovery was 64% (+/-26%), and in 25/35 (71%) harvesting procedures the minimum progenitor number required for safe autografting could be obtained by one single leukapheresis. We conclude that the Fresenius AS 104 blood cell separator provides a high cell yield and is a safe device for leukapheresis in pediatric patients.     

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.     

Optimizing pediatric peripheral blood stem cell collection

IntroductionPediatric PBSC harvests pose specific challenges during apheresis and a knowledge of the same and variables affecting PBSC collection are very important in planning these procedures. In the present study safety profile of pediatric PBSC procedures and variables influencing the successful collection were analyzed.MethodPediatric PBSC harvest data for 3 years was reviewed for donor, procedural and product parameters and any specific challenges faced during the procedures. Successful PBSC collection was defined when CD34 dose obtained was ≥2 × 10⁶ cells/Kg of recipients’ body weight.Results85 PBSC collections performed on 46 children (age range 1.5–15 years) were included. Sixty-two procedures were on autologous donors and 23 on allogenic donors. The median CD34+ cell dose in the PBSC product per procedure was 2.12 × 10⁶ cells/Kg for autologous procedures and 4.6 × 10⁶ cells/Kg for allogenic procedures. Systemic adverse reaction was observed during only one procedure (0.01 %) and was managed conservatively. Successful dose was collected in 52 procedures (61.17 %) and was significantly associated with CD34+ count of more than 19.7/μL, monocyte count of more than 1.65 × 10⁶/μL, allogenic collection and female gender (p = 0.00001, p = 0.011, p = 0.00052, and p = 0.0001, respectively).ConclusionPBSC collection is safe in pediatric age groups and pre-procedure CD34 count of ≥20/μL on the day of collection may result in successful collection of stem cell dose. It is important to identify factors associated with failed collection for appropriate counselling and justifying pre-emptive use of stem cell mobilizing agents.     

Efficacy of peripheral blood stem cell transplantation

With the advancement in apheresis technique and collection, the role of peripheral blood stem cell (PBSC) transplantation has emerged. PBSC are now being utilized either alone to reconstitute hematopoiesis following high-dose myeloablative therapy, or in combination with autologous bone marrow transplantation to enhance hematopoietic recovery, or as supportive therapy to allow dose-intensity of conventional chemotherapy. The role and efficacy of PBSC transplantation will be reviewed in the article.     

外周血干细胞动员研究现状

外周血干细胞(PBSC)因其获取便利,已广泛应用于临床造血干细胞移植.迄今已有大量研究阐明造血干细胞(HSC)骨髓定植机制,以及如何通过扰乱这种机制促使HSC释放于外周血中,并已发现用于PBSC动员的新一代药物--普乐沙福(Plerixafer)与甲状旁腺素(PTH)等联合G-CSF用于PBSC动员,大大提高了动员效率,其中Plerixafer已被FDA批准用于临床.     

Thrombotic microangiopathy during peripheral blood stem cell mobilization

Granulocyte colony‐stimulating factor (GCSF) is currently the most widely used cytokine for stem cell mobilization. There are few studies suggesting GCSF administration may induce activation of both coagulation and endothelial cells that could favor the developing of thrombotic events. We report a 58‐year‐old female with vasculitis and renal impairment. She was found to have an underlying monoclonal gammopathy of unknown significance (MGUS). The monoclonal protein was felt to play a role in her underlying renal disease and peripheral neuropathy. She was considered a candidate for peripheral blood stem cell transplantation to manage the monoclonal protein. During stem cell mobilization with GCSF, she developed worsening of anemia; thrombocytopenia and worsening of renal function. She was diagnosed with thrombotic microangiopathy (TMA) which was successfully treated with therapeutic plasma exchange and rituximab. It is possible that GCSF may have directly (activating endothelial cells) or indirectly (activation of underlying autoimmune disorder) contributed to TMA in this patient. J. Clin. Apheresis 2009. © 2009 Wiley‐Liss, Inc.     

Haploidentical stem cell transplantation-bone marrow vs peripheral blood

Hematopoietic stem cells may be obtained by collection of bone marrow, mobilization and collection of peripheral blood stem cells or umbilical cord blood. Transplantation of peripheral blood hematopoietic cells has increased due to faster engraftment and practicability in both the related, unrelated or haploidentical setting. We reviewed the question of which stem cell source - bone marrow (BM) or peripheral blood (PBSC) - is the most suitable for individuals undergoing haploidentical stem cell transplantation. BM or PBSC could be safely used as allograft sources for haploidentical transplantation with good outcomes and acceptable rates of GVHD and graft failure. Prospective randomized studies are needed to evaluate the effect of PB vs BM in haploidentical settings.     

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.     

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。     

Treatment for the decline of ionized calcium levels during peripheral blood progenitor cell harvesting

BACKGROUND: ACD-A solution containing sodium citrate and citric acid is used as an anticoagulant agent during peripheral blood progenitor cell (PBPC) harvesting, and in rare cases can cause fatal citrate intoxication. The aim of this study was to establish effective methods for stabilizing ionized calcium (ICa) levels during PBPC harvesting. STUDY DESIGN AND METHODS: ICa was measured during 46 apheresis procedures conducted in 26 patients. Four patients in four procedures were infused with calcium gluconate solution before PBPC harvesting; three patients in six procedures were infused with calcium gluconate when symptoms of citrate intoxication appeared; and four patients in five procedures received a continuous infusion. Five patients in five procedures took an isotonic sports drink containing calcium when hypocalcemic symptoms appeared. The ICa level, blood pressure, and pulse rate were measured. RESULTS: ICa declined rapidly from the preapheresis level of 1.081(+/-0.092) mM to 0.937(+/-0.081) mM (13.3%, p < 0.0001) 10 minutes after the start of apheresis and continued to decline until the completion of the procedure. When patients received a continuous infusion of calcium during apheresis, ICa was relatively stabilized. ICa significantly rose (6.1 +/- 3.6%, p < 0.02) within 2 to 5 minutes after oral intake of an isotonic sports drink containing calcium and was maintained within normal range for 31 to 55 minutes. CONCLUSION: An isotonic sports drink containing calcium has a quick stabilizing and a longer maintenance effect on ICa. Thus, we recommend the intake of an isotonic sports drink containing calcium as the easiest and best method for preventing hypocalcemia during apheresis.     

Nonmyeloablative allogeneic peripheral blood stem cell transplantation of renal cell carcinoma

We presented three cases with refractory metastatic renal cell carcinomas who underwent nonmyeloablative allogeneic peripheral blood stem cell transplantation(PBSCT). A fludarabine and cyclophosphamide-based regimen was used. Complete donor-T-cell chimerism was established at day 94, 40, 52 after PBSCT in each case. GVHD occurred at day 183, 53, 75. In Case 1 and Case 3 apparent GVT effects reduced the metastases at day 212, 261 and let them live longer than expected. Case 2 died at day 69 before GVT effect appeared, because the case was so-called rapid growth type. Histological examinations in Case 1 and Case 3 showed infiltration of lymphocytes disrupting the foci of renal cell carcinomas which was considered as findings of GVT effects.     

自体外周血干细胞移植治疗周围神经损伤

背景:关于神经干细胞对周围神经损伤的治疗已有多篇报道,但外周血干细胞对周围神经损伤治疗鲜有报道。目的:探讨自体外周血干细胞移植治疗周围神经损伤使失神经骨骼肌重获神经再支配的临床应用。方法:应用外周血干细胞治疗周围神经损伤6例,同时与周围神经损伤单纯行神经断端吻合或神经移植10例比较。2组患者术后常规肌注鼠神经生长因子一两个疗程,同时给予针灸、理疗、经皮电刺激治疗及功能康复训练。结果与结论:两组患者随访均超过6个月。干细胞移植组运动神经传导速度和感觉神经传导速度的恢复率要明显高于单纯神经吻合组。提示周围神经损伤后给予修复局部用外周血干细胞移植能够使远端失神经骨骼肌早期重新获得神经再支配。