Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources.

Endothelial progenitor cells (EPCs) have been broadly studied for several years due to their outstanding regenerative potential. Moreover, these cells might be a valuable source of genetic information for the preservation of endangered animal species. However, a controversy regarding their characterization still exists. The aim of this study was to isolate and compare the rabbit peripheral blood- and bone marrow-derived EPCs with human umbilical vein endothelial cells (HUVECs) in terms of their phenotype and morphology that could be affected by the passage number or cryopreservation as well as to assess their possible neuro-differentiation potential. Briefly, cells were isolated and cultured under standard endothelial conditions until passage 3. The morphological changes during the culture were monitored and each passage was analyzed for the typical phenotype using flow cytometry, quantitative real-time polymerase chain reaction (qPCR) and novel digital droplet PCR (ddPCR), and compared to HUVECs. The neurogenic differentiation was induced using a commercial kit. Rabbit cells were also cryopreserved for at least 3 months and then analyzed after thawing. According to the obtained results, both rabbit EPCs exhibit a spindle-shaped morphology and high proliferation rate. The both cell lines possess same stable phenotype: CD14-CD29+CD31-CD34-CD44+CD45-CD49f+CD73+CD90+CD105+CD133-CD146-CD166+VE-cadherin+VEGFR-2+SSEA-4+MSCA-1-vWF+eNOS+AcLDL+ALDH+vimentin+desmin+α-SMA+, slightly different from HUVECs. Moreover, both induced rabbit EPCs exhibit neuron-like morphological changes and expression of neuronal markers ENO2 and MAP2. In addition, cryopreserved rabbit cells maintained high viability (>85%) and endothelial phenotype after thawing. In conclusion, our findings suggest that cells expanded from the rabbit peripheral blood and bone marrow are of the endothelial origin with a stable marker expression and interesting proliferation and differentiation capacity.

Influence of laboratory procedures on postthawing cell viability and hematopoietic engraftment after autologous peripheral blood stem cell transplantation.

BACKGROUND: The kinetics of hematopoietic recovery after autologous stem cell transplantation (ASCT) may be affected by laboratory procedures. The aim of this study was to evaluate the influence of characteristics of the cryopreserved units of peripheral blood stem cells (PBSC) on postthawing cell viability and engraftment outcomes after ASCT. STUDY DESIGN AND METHODS: This was a retrospective cohort study including individuals referred for ASCT. Cryopreservation was conducted at a single processing facility between 2014 and 2019, and patients received clinical care at six transplant centers. Covariates and outcome data were retrieved from participants' records. RESULTS: The study population comprised 619 patients (345 [55.7%] male). Median age was 53 years. Multiple myeloma was the most common diagnosis (62.7%). Higher preapheresis CD34+ cell count, lower nucleated cell (NC) concentration per cryobag, and composition of the cryoprotectant solution (5% dimethyl sulfoxide [DMSO] and 6% hydroxyethyl starch) were statistically significantly associated with higher postthawing cell viability. The linear regression model for time to neutrophil and platelet engraftment included the infused CD34+ cell dose and the composition of the cryoprotectant solution. Patients who had PBSC cryopreserved using 10% DMSO solution presented six times higher odds (odds ratio [OR] = 6.9; 95% confidence interval [CI]: 2.2-21.1; p = .001) of delayed neutrophil engraftment (>14 days) and two times higher odds (OR = 2.3, 95%CI: 1.4-3.7; p = .001) of prolonged hospitalization (>18 days). DISCUSSION: The study showed that mobilization efficacy, NC concentration, and the composition of the cryoprotectant solution significantly affected postthawing cell viability. In addition, the composition of the cryoprotectant solution significantly impacted engraftment outcomes and time of hospitalization after ASCT.

Comparison of spectra optia and amicus cell separators for autologous peripheral blood stem cell collection.

INTRODUCTION: Autologous peripheral blood stem cell (PBSC) transplantation has become a standard treatment option for many oncology patients. The aim of this study was to evaluate the performance of two cell separators, Spectra Optia (Terumo BCT, Japan) and Amicus (Fresenius-Kabi) for autologous PBSC collection. METHODS: We retrospectively evaluated 56 apheresis by Spectra Optia with Continuous Mononuclear Cell Collection (cMNC) from 20 patients, and 50 apheresis by Amicus from 27 patients between December 2018 and December 2019. CD34+ collection efficiency (CE2) and platelet (PLT) loss were evaluated. RESULTS: There was no significant difference in CD34+ CE2 between Spectra Optia with cMNC (median, 28.8%) and Amicus (median, 33.1%; P = 0.537). PLT loss was significantly lower in Amicus (median, 28.6%) than in Spectra Optia with cMNC (median, 37.8%; P = 0.009). CONCLUSION: CD34+ CE2 was comparable between Spectra Optia and Amicus, and PLT loss was significantly lower in Amicus. To the best of our knowledge, this is the first report comparing autologous PBSC collection of the Spectra Optia and Amicus. These results may provide general guidance with regard to device selection to apheresis clinics that use both separators for optimal outcomes depending on each patient's characteristics.

Peripheral blood stem cell collection in children with extremely low body weight (≤8 kg). What have we learned over the past 25 years and where are the limits?

Hematopoietic progenitor cells-apheresis (HPC-A) collection is now a routine procedure for autologous hematopoietic stem cell transplantation. Here we present our 25 years' experience of HPC-A collection in children weighing 8 kg or less, with a focus on the evolution of our standard operating procedures, and the safety limits for these young patients, in the Pediatric Apheresis Unit of Clermont-Ferrand University Hospital (France). Fifteen children weighing 8 kg or less underwent 26 HPC-A collections over 25 years. Median CD34+ cell yield by leukapheresis was 4.4 106 /kg. No procedure-related complications were encountered during or after the collection. No patient had profound thrombocytopenia or anemia that needed post-collection transfusions. Our experience in pediatric oncology patients who underwent HPC-A collections shows that this procedure can be performed even in the smallest of children with no increase in toxicity provided all precautions are taken to ensure that the procedure is carried out under the ideal conditions.

Generation of a Human Allergic Mast Cell Phenotype from CD133+ Stem Cells.

Cultured human mast cells are a useful tool for research into innate immune responses as well as allergic mechanisms. Mast cells cultured from peripheral blood can provide information on immune mechanisms of known, selected individuals. With the method presented here, eight million mast cells can be cultured from ca. one million stem cells purified from one unit (450 mL) of human peripheral blood. Culture with IgE and IL4 optimizes an allergic phenotype of the mast cells.

Generation and Culture of Peripheral CD34+ Stem Cell-Derived Mast Cells (PSCMCs).

Mast cells (MCs) are long-living tissue-resident cells that play an important role in inflammatory and allergic reactions. In vitro models of mast cell functions have allowed better understanding of the function of mast cells and mast cell-mediated disorders. In this unit, we describe a protocol used for the generation and culture of peripheral CD34+ stem cell-derived mast cells (PSCMCs). It provides a useful tool for the investigation of the biology of human MCs in vitro.

Modulating the Human Basophil Phenotype During Its Development and Maturation: Basophils Derived from In Vitro Cultures of CD34+ Progenitor Cells.

Historically, the human basophil that is studied experimentally comes from peripheral blood. But there is evidence that only a short portion of the basophil life cycle related to IgE-mediated function occurs in the blood. The same evidence suggests that IgE-mediated functionality is present for 5-7 days in the bone marrow (or other tissues) during which the cell modulates its phenotype according to local conditions. It is suggested that to properly understand the nature of basophil behavior, a better understanding of its biology during maturation would be helpful. For example, one highly suggestive line of evidence for the relevance of understanding the maturation period is related to the change in basophil phenotype that occurs during treatment of patients with omalizumab. During this treatment, the intrinsic reactivity or sensitivity of the basophils is significantly increased despite, or perhaps because of, the dramatic reduction in FcεRI expression that accompanies this treatment. One of the critical signaling enzymes to increase expression selectively in basophils during treatment is SYK, which is one of the earliest signaling tyrosine kinases involved in translating the aggregation of FcεRI into secretion from the cell. Treatment with omalizumab increases SYK expression, and this observation focuses some attention of how SYK expression is regulated. It is possible that the key regulation occurs during maturation of the basophil. Regardless of the mechanisms operative in this particular treatment, understanding the process of maturation and the extrinsic factors that influence it may lead to better understanding of disease processes. Therefore, this chapter will discuss and present techniques to work with maturing human basophils.

Review of conditioning regimens for haplo-identical donor transplants using post-transplant cyclophosphamide in recipients of G-CSF mobilised peripheral stem cell.

Haplo-identical transplant is being increasingly used in patients who do not have a readily available matched related or unrelated donor. Post-transplant cyclophosphamide's use due to its simplicity and documented efficacy has made this approach readily employable across diverse transplant centres across the globe. The outcomes of regimens used for conditioning in recipients of bone marrow are at times in variance to that from more commonly employed G-CSF mobilised peripheral stem cell (PBSC). This review highlights various conditioning regimens used in PBSC recipients, with emphasis on toxicities, practicalities and transplant related outcomes of relapse, non-relapse mortality and graft versus host disease.

Assessment and streamlined preparation of low-cytotoxicity lentiviral vectors for mobilized human hematopoietic stem cell transduction.

As important vectors for ectopic protein expression, gene silencing, and progenitor cell barcoding, lentiviruses continue to emerge as versatile research and clinical tools. For studies employing cell types that are relatively resistant to transduction, high-titer lentivirus preparations with low cytotoxicity are required. During lentivirus production, carryover plasmid DNA endotoxins, transfection reagents, damaged packaging cells, and virus concentration procedures are potential sources of cytotoxicity. As an often unevaluated property of lentivirus preparations, cytotoxicity can unwittingly skew estimates of functional titers and complicate interpretations of transduced cell phenotypes. By employing hematopoietic UT7epo cells cultured in erythropoietin (EPO) below maximal dosing, we first define a sensitive flow cytometric bioassay for critically assessing the cytotoxicity (and titers) of lentivirus preparations. Bioassay of custom preparations of research-grade lentiviruses from six commercial sources unexpectedly revealed substantial cytotoxicity (with certain preparations additionally registering titers several log below designated values). To overcome such limiting properties, we further report on unique, efficient workflows for reproducibly preparing and processing high-titer, low-cytotoxicity (HTLC) lentiviruses at research scale. These HTLC lentiviruses reliably transduce peripheral blood hematopoietic stem/progenitor cells (PB-HSPCs) at frequencies ≥40%, with low cytotoxicity. In addition, by employing cyclosporin H (to inhibit IFITM3), PB-HSPCs can be transduced at heightened efficiency with nominal cytotoxicity. Overall, this work provides straightforward approaches to (1) critical assessment of the cytotoxicity of lentivirus preparations; (2) reproducible generation (and concentration) of high-quality lentiviruses via a streamlined workflow; and (3) transduction of PB-HSPCs at benchmark levels with nominal cytotoxicity.

Apheresis machines variably overestimate mononuclear cell collection volume.

INTRODUCTION: Calculation of the actual number of CD34+ cells in the collection product is based on the volume of the collected product and its concentration of CD34+ cells measured in the lab. The number of CD34+ cells infused correlates closely with the pace of hematopoietic reconstitution following autologous or allogeneic stem cell transplantation. METHODS: We studied peripheral blood stem cell collections in a single apheresis center with two Spectra Optia devices, using mononuclear cell collection or continuous mononuclear cell collection procedures. The collection volume displayed by the apheresis device was compared with the volume determined by a weight-based method. RESULTS: Fifty-two consecutive CD34 collections in 35 different donors (range 1-4 daily procedures per donor) were analyzed. The machines reported larger collection volumes (P < .001). The mean collection volume reported by the machine was 274.37 mL (range 162-396). The mean manually measured collection volume was 261.82 mL (range 155-371.40). Mean overestimation by machine was 12.53 mL (range -0.95 to 31.24; 95% confidence interval 10.94-14.11) or 4.88% (range -0.26 to 10.28). Median overestimation of the absolute number of CD34 was 10.29 × 106 (range -2.83 to 141.84 × 106 ). CONCLUSION: Both Spectra Optia machines overestimated the collection volume after peripheral blood stem cell collection. Although the mean variation falls within the expected range, in some cases, this overestimation may be clinically relevant if no other method of measurement is used.

Low-dose peripheral blood stem cell graft after high-dose chemotherapy - an evaluation of hematopoietic reconstitution.

BACKGROUND: High-dose (HD) chemotherapy followed by autologous blood stem-cell transplantation (ASCT) is the standard treatment for multiple myeloma (MM) patients. However, the collection of sufficient peripheral blood stem cell (PBSC) grafts can be challenging, and the question arises whether reinfusion of low-dose grafts will lead to a hematopoietic recovery. METHODS: The hematopoietic recovery of 148 MM patients who underwent HD melphalan chemotherapy and received PBSC transplants with varying CD34+ cells doses (3-4 × 106 [n = 86], 2-2.5 × 106 [n = 53], < 2 × 106 [n = 9] per kg body weight [bw]) was analyzed in this retrospective single-center study. RESULTS: All patients reached hematopoietic reconstitution, even those who received < 2 × 106 CD34+ cells/kg bw. 62 (42%) patients received granulocyte-colony-stimulating factor (G-CSF). The median duration to leukocyte recovery ≥1.0 × 109/L was 12 days in every group. The median duration to platelet recovery ≥20 × 109/L was 11, 13 and 13 days, respectively. In the multivariate analysis, a low number of reinfused CD34+ cells was associated with prolonged time until leukocyte reconstitution (p = 0.010, HR 0.607) and platelet recovery (p < 0.001, HR 0.438). G-CSF support significantly accelerated leukocyte (p < 0.001, HR 16.742) but not platelet reconstitution. CONCLUSION: In conclusion, reinfusion of low- and even very-low-dose PBSC grafts leads to sufficient hematopoietic reconstitution. No severe adverse events were observed during or after HD chemotherapy and ASCT in the analyzed cohort. While the impact of CD34+ cell dose is marginal, G-CSF significantly accelerates the leukocyte recovery.

A new protocol for improving efficiency of autologous peripheral blood stem cell collection in patients with high white blood cell counts.

BACKGROUND: Collection efficiency (CE) of peripheral blood stem cell (PBSC) collections is negatively affected by increasing white blood cell (WBC) counts of the patient. This study compared a new optimized mononuclear cell (MNC) collection protocol (OPP) to the standard MNC collection protocol recommended by the manufacturer (STP) for PBSC collection in patients with WBC counts >35 000/µL. STUDY DESIGN AND METHODS: Single-center, retrospective, and observational study of 81 autologous PBSC collections on Fenwal Amicus cell separators in 70 adult patients. RESULTS: Median peripheral WBC count (×103 /µL; 44.2 in OPP group vs 46.5 in STP group) and median CD34+ count (105/µL in OPP group vs 40/µL in STP group) at the beginning of PBSC collection did not differ significantly. Median CE2 (45% vs 31%; P < .001) as well as CD34+ yield of the apheresis product both with regards to median absolute CD34+ content (×106 ; 793 vs 188; P = .001) as well as median CD34+ content (×106 )/kg body weight (8.93 vs 2.51; P = .002) were significantly higher for the OPP. Overall, 18/21 (86%) patients with the OPP obtained their target CD34+ amount with a single apheresis session, compared to 25/50 (50%) with the STP (P = .005). PBSC collections using OPP lasted significantly longer (median 377 minutes vs 260 minutes; P < .001) than with the STP. CONCLUSIONS: The OPP significantly improves CE2 for PBSC collections on Fenwal Amicus cell separators in patients with pre-apheresis WBC counts >35 000/µL and significantly reduces the necessity for multiple apheresis sessions. The OPP is therefore suited to reduce both patient burden and cost in autologous PBSC collection.

Successful Stem Cell Apheresis Using Spectra Optia in a 6 kg Child With Atypical Teratoid/Rhabdoid Tumor.

Peripheral blood stem cell apheresis has become a routine procedure for the collection of peripheral blood stem cells to enable high-dose chemotherapy followed by autologous stem cell transplantation in high-risk pediatric malignancies. However, the procedure remains challenging in very low-weight infants due to high extracorporeal blood volume and citrate toxicity. Our case report demonstrates in detail a successful and complication-free large-volume leukapheresis in a very small infant weighing 6 kg using a Spectra Optia apheresis system after placing a femoral double-lumen Shaldon catheter. Anticoagulation was achieved by citrate dextrose solution without the use of heparin. The total amount of blood being processed during the procedure equaled almost 4 times the total blood volume of the patient. The final apheresis product contained 14.0×10 CD34 cells/kg body weight. The infant was diagnosed with an atypical teratoid/rhabdoid tumor of the thalamus and third ventricle at the age of 3 months and had a history of epileptic seizures.

Assessment of haematopoietic progenitor cell counting with the Sysmex® XN-1000 to guide timing of apheresis of peripheral blood stem cells.

BACKGROUND: Successful peripheral blood stem cell (PBSC) collection depends on optimal timing of apheresis, as usually determined by flow cytometry CD34-positive (+) cell count in peripheral blood (PB). Since this method is costly and labour-intensive, we evaluated the use of the Hematopoietic Progenitor Cell count programme on a Sysmex® XN haematologic analyser (XN-HPC) as a rapid and inexpensive alternative for predicting CD34+ cell count in PB samples. MATERIALS AND METHODS: Haematopoietic progenitor cell and CD34+ cell counts were compared using 273 PB samples collected from 78 healthy donors and 72 patients who underwent PBSC transplantation. We assessed the effectiveness of the XN-HPC in safely predicting pre-harvest CD34+ counts. The most efficient cut-off values of XNHPC were identified. We also evaluated the imprecision (coefficient of variation, CV) and functional sensitivity. RESULTS: Imprecision of the XN-HPC count was <6.3% on daily measurement of three levels of quality control material. Functional sensitivity was 8.9×106/L. A cut-off value of ≥62×106/L XN-HPC for multiple myeloma (MM) patients and ≥30×106/L for all other subjects had both 100% specificity and 100% positive predictive value for identifying samples with CD34+ cells ≥20×106/L. An XN-HPC threshold of <13×106/L identified preharvest CD34+ cell count <10×106/L with 100% sensitivity and 100% negative predictive value. DISCUSSION: The XN-HPC is a fast, easy and inexpensive test that can safely improve apheresis workflow thus possibly replacing other more expensive CD34 counts currently performed and promoting optimal timing of PBSC collection.

The comparison of effectiveness and safety between different biosimilars of G-CSF in the mobilization of peripheral blood stem cells (PBSCs) for autologous transplantation (autologous peripheral blood stem cell transplantation, auto-PBSCT).

BACKGROUND: Autologous peripheral blood marrow stem cell transplantation (auto-PBSCT) preceded by high-dose chemotherapy is a well-known method of treatment for patients with hematological cancers. Performing the procedure entails obtaining from the patient their own stem cells from peripheral blood using G-CSF. Currently, various filgrastim biosimilars are widely used. AIM OF THE STUDY: The purpose of this study is to compare the efficacy and safety of three different biosimilars of filgrastim in PBSC mobilization in patients with hematological malignancies. MATERIALS AND METHODS: This is a retrospective analysis of 282 patients (118 women and 164 men) who underwent stem cells mobilization for auto-PBSCT in the Department of Hematology in Wroclaw in 2012-2014. Three filgrastim biosimilars were used: Tevagrastim (95), Nivestim (92), and Zarzio (95). Ninety patients (32%) were diagnosed with multiple myeloma, 55 (19%) with Hodgkin's lymphoma, 90 (32%) with NHLs, 20 (7%) with acute myeloid leukemia, and 27 (10%) with another hematological cancer. RESULTS: The mean number of CD34+ cells collected during the first leukapheresis was 5.95 × 106 /kg for Tevagrastim, 7.08 × 106 /kg for Nivestim, and 6.8 × 106 /kg for Zarzio (P > .05). The necessary number of leukapheresis for patients receiving Zarzio, Nivestim, and Tevagrastim was 1.32, 1.37, and 1.66, respectively (P > .05). The percentage of effective mobilizations was 88.2% for Zarzio, 86.2% for Nivestim, and 84.9% for Tevagrastim. The side effects included bone pain and headache. CONCLUSION: All tested biosimilars demonstrated similar effectiveness and safety profiles in patients with hematological tumors undergoing PBSC mobilization; therefore, they can be used interchangeably.

Smoking is an important factor that affects peripheral blood progenitor cells yield in healthy male donors.

BACKGROUND: Smoking could reduce the CD34+ cells in peripheral blood of healthy individual. This study aimed to investigate the correlation between smoking load and the effect of peripheral blood hematopoietic progenitor cells (PBPCs) mobilization by granulocyte colony-stimulating factor (G-CSF) alone in healthy donors. METHODS: Retrospective analysis was performed on 145 healthy adult PBPCs donors who underwent PBPCs mobilization and collection. Smoking factors were evaluated and correlated with mobilization responses, as indicated by the collected CD34+ cells concentration. RESULTS: The collected CD34+ cells concentration was closely related to pre-CD34 (P < .001) and CD34+ cells collected per volume blood processed (P < .001) which suggested that collected CD34+ cells concentration was a reliable indicator of PBPCs mobilization efficiency. The heavy smoking donors revealed significantly lower collected CD34+ cells concentration, compared to that of the nonsmoking (P < .001) and light smoking donors (P < .05). The levels of collected CD34+ cells in light smoking were also obviously lower than that in nonsmoking donors (P < .05).There were no obvious differences in the collected CD34+ cells concentration, overall processed blood volume and total collected CD34+ cells between nonsmoking and smoking cessation groups (P = .490; P = .464; P = .819). CONCLUSION: Cigarette smoking is an important factor that affects the yield of PBPCs in male donors, especially when the smoking load is more than five pack-years. Mobilization of PBMCs could be restored by smoking cessation in chronic smokers.

Derivation of a MUSIi012-A iPSCs from mobilized peripheral blood stem cells.

CD34+ cells were isolated from mobilized peripheral blood of a healthy donor and reprogrammed by nucleofection with episomal plasmids carrying l-MYC, LIN28, OCT4, SOX2, KLF4, EBNA-1, and shRNA against p53. The obtained MUSIi012-A cell line maintained the pluripotent phenotype, the ability to differentiate into all three germ layers, and a normal karyotype.

Impact of CD34+ pre-counting and plerixafor on autologous peripheral blood stem cell collection in Japanese university hospitals in eight years.

OBJECTIVE: Over the past decade, there have been two major advancements in autologous peripheral blood stem cell (PBSC) collection, namely enumeration of CD 34+ cells for apheresis prediction and use of plerixafor to assist mobilization of PBSC. This study aimed to investigate changes in the efficacy of PBSC collection from two Japanese university hospitals over an eight-year period. STUDY DESIGN AND METHODS: A series of 399 PBSC collection procedures from 239 patients with solid malignant tumors (ST, n = 42), malignant lymphoma (ML, n = 91), multiple myeloma (MM, n = 99), and others (amyloidosis and leukemia, n = 7) from two university hospitals from 2011 to 2018 were retrospectively analyzed. We also analyzed the effects of CD34+ pre-counting and plerixafor administration in improving CD34+ cell yield. RESULTS: Using CD34+ pre-count as a reference, the frequency of apheresis was reduced and the yield of CD34+ cells increased in patients with ST. When administrating plerixafor, especially with a CD34+ pre-count <20/µL, the yield of CD34+ cells was significantly increased in patients with ML (p = 0.02) and MM (p = 0.03). CONCLUSIONS: We verified that CD34+ cell counting and plerixafor administration contributed to effective PBSC collections in our hospitals for the eight-year study period. In patients with ST, CD34+ pre-count threshold for starting apheresis was ≥10/µL. CD34+ pre-count (<20/µL) was useful to select appropriate patients for plerixafor administration among the patients with ML and MM.

A matched-pair analysis reveals marginally reduced CD34+ cell mobilization on second occasion in 27 related donors who underwent peripheral blood stem cell collection twice at the same institution.

BACKGROUND: In a small proportion of cases, hematopoietic function is insufficient after allogeneic hematopoietic stem cell transplantation, as a result of poor graft function or graft failure. These complications are common indications of re-mobilization of the initial donor, either for a second allograft or for an infusion of CD34+ Selected stem Cell Boost (SCB). METHODS AND MATERIALS: We retrospectively reviewed the results of two cycles of CD34+ cell mobilization and collection. CD34+ cells mobilized and collected at each cycle were compared. When CD34+ cell selection from the collected allogeneic mononuclear cells was indicated, it was performed with the Clinimacs Plus® medical device, and results from in-process and final quality checks were analyzed. To assess the efficacy of CD34+ SCB, transfusion needs before and after the infusion of selected CD34+ cells were calculated. RESULTS: The median peripheral blood concentration of CD34+ cells/µL was marginally reduced during the second cycle (35.6 vs 33.8, p < 0.05); results revealed a strong correlation between paired values (r = 0.85). The cumulative number of collected CD34+ cells were similar for both cycles; the total processed blood volume was higher during the second cycle (p = 0.023). For CD34+ immune-selection procedures, CD34+ cell recovery and purity were respectively 57% and 95%, with a median T-cell depletion of 6.7 log. Recipients' needs for platelet and red blood cell transfusions were significantly reduced after CD34+ SCB. CONCLUSION: This study confirms the feasibility of a second cycle of mobilization in healthy related donors and the benefits of CD34+ SCB on hematopoietic reconstitution.