Quantification of peripheral blood CD34+ cells prior to stem cell harvesting by leukapheresis: a single center experience

Lemos, Natália Emerim; Farias, Mariela Granero; Kubaski, Francyne; Scotti, Luciana; Onsten, Tor Gunnar Hugo; Brondani, Letícia de Almeida; Wagner, Sandrine Comparsi; Sekine, Leo

doi:10.1016/j.htct.2018.01.002

Cited by 16 publications

(13 citation statements)

References 24 publications

(45 reference statements)

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“…However, the pre-harvest concentration and post-harvest cell dose were directly proportional to each other. This was also seen in a study by Lemos et al where they have highlighted a moderate positive correlation between peripheral blood CD34+ cell count and total CD34+ cell count/kg (r=0.596; p <0.001) 29 .…”

Section: Discussionsupporting

confidence: 75%

Pattern of autologous stem cell transplants at a tertiary care government hospital, with emphasis on transplant outcomes with pre-harvest CD34+ level

Dolai

Sen

et al. 2022

BCT

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Purpose: Autologous stem cell transplantation (ASCT) is an established therapy for many hematological diseases. This study assessed the pattern of ASCTs at a tertiary care center and associated factors, including pre-harvest CD34+ stem cell levels, leading to improved engraftment outcomes. Methodology: A retrospective study was conducted in India, between February 2009-August 2020. Patients who underwent ASCT for different hematological malignancies (n=65) were included, and the patients' age, sex, type and stage of disease, pre-and post-harvest CD34+ counts, and time to attain platelet/neutrophil engraftment or febrile neutropenia were analyzed. The post-harvest CD34+ dose was calculated. Pre-conditioning was performed using Granulocyte Colony Stimulating Factor (GCSF)±Plerixafor. Progression-free survival (PFS) was calculated using relapse/death as the endpoint. Results:The median age of the cohort (n=65) was 49 years, with a male preponderance. Multiple myeloma was the most common malignancy (70.8% [46/65]), requiring ASCT. The median time to ASCT was 13 months. All patients had received GCSF, while Plerixafor was used in 17 patients with a pre-harvest CD34+ count of <10 cells/ μL. The median pre-harvest CD34+ concentration and post-harvest CD34+ cell dose was 27.54 cells/μL (n=26) and 5.23×10 6 cells/kg body weight (n=65), respectively. The median time to engraftment was 11 and 12 days, for neutrophils and platelets, respectively. One patient did not engraft and was excluded from the analysis. The time required to attain neutrophil engraftment was significantly lower (p=0.02) among freshly harvested stem cells (n=48) than that of cryopreserved products (n=17). Platelet engraftment associated with CD34+ pre-and post-harvest levels was not significant (p=0.06). The time to attain neutropenia and subsequent febrile neutropenia was significantly lower with an adequate post-harvest CD34+ dose (p=0.009). Febrile neutropenia was seen in 83.1% (54/65) patients. The median time for febrile neutropenia was 4 days post-ASCT. Pre-and postharvest CD34+ concentrations were directly proportional to each other (p<0.001). The median PFS was 112 months (n=65). Survival was better in males (median PFS: 112 months) vs. females (median PFS: 59 months) (p= 0.27). Eight patients relapsed, and eight patients had died. Conclusion: Although unrelated to age or sex, the post-harvest CD34+ dose was inversely related to febrile neutropenia. As pre-and post-harvest CD34+ levels were directly proportional, pre-harvest CD34+ concentrations may be reliably used to assess engraftment outcomes. Rapid neutrophil engraftment was noted in fresh stem cells with PFS of 112 months, and was better among males, the exact reason being unknown. Thus, a larger number of patients should be followed up to obtain an accurate picture.

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Section: Discussionsupporting

confidence: 75%

Pattern of autologous stem cell transplants at a tertiary care government hospital, with emphasis on transplant outcomes with pre-harvest CD34+ level

Dolai

Sen

et al. 2022

BCT

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“…Surface antigens were quantified by flow cytometry using ZE5 Cell Analyzer (Bio-Rad) and BD FACSAria™ III (BD). PE-labeled mouse anti-human CD34 [ 27 ], FITC-labeled mouse anti-mouse CD45.1, FITC-labeled mouse anti-NHP CD45, V450 mouse lineage antibody cocktail with isotype control, PE-labeled mouse anti-mouse CD45.2, FITC-labeled hamster anti-mouse CD48, PE-Cy™7-labeled rat anti-mouse Ly-6A/E, APC-labeled rat anti-mouse CD117 (BD Biosciences), and PE-labeled anti-mouse CD150 (SLAM) (BioLegend) antibodies were used in the flow cytometry assays.…”

Section: Methodsmentioning

confidence: 99%

A novel small molecule CXCR4 antagonist potently mobilizes hematopoietic stem cells in mice and monkeys

et al. 2021

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Background Hematopoietic stem cell (HSC) transplantation is an effective treatment strategy for many types of diseases. Peripheral blood (PB) is the most commonly used source of bone marrow (BM)-derived stem cells for current HSC transplantation. However, PB usually contains very few HSCs under normal conditions, as these cells are normally retained within the BM. This retention depends on the interaction between the CXC chemokine receptor 4 (CXCR4) expressed on the HSCs and its natural chemokine ligand, stromal cell-derived factor (SDF)-1α (also named CXCL12) present in the BM stromal microenvironment. In clinical practice, blocking this interaction with a CXCR4 antagonist can induce the rapid mobilization of HSCs from the BM into the PB. Methods C3H/HEJ, DBA/2, CD45.1+, and CD45.2+ mice and monkeys were employed in colony-forming unit (CFU) assays, flow cytometry assays, and competitive/noncompetitive transplantation assays, to assess the short-term mobilization efficacy of HF51116 and the long-term repopulating (LTR) ability of HSCs. Kinetics of different blood cells and the concentration of HF51116 in PB were also explored by blood routine examinations and pharmacokinetic assays. Results In this paper, we report that a novel small molecule CXCR4 antagonist, HF51116, which was designed and synthesized by our laboratory, can rapidly and potently mobilize HSCs from BM to PB in mice and monkeys. HF51116 not only mobilized HSCs when used alone but also synergized with the mobilizing effects of granulocyte colony-stimulating factor (G-CSF) after co-administration. Following mobilization by HF51116 and G-CSF, the long-term repopulating (LTR) and self-renewing HSCs were sufficiently engrafted in primary and secondary lethally irradiated mice and were able to rescue and support long-term mouse survival. In monkeys, HF51116 exhibited strong HSC mobilization activity and quickly reached the highest in vivo blood drug concentration. Conclusions These results demonstrate that HF51116 is a new promising stem cell mobilizer which specifically targets CXCR4 and merits further preclinical and clinical studies.

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“…Mesenchymal [33] and hematopoietic [34] stem cells will be obtained from nine children with indication of blood panel exam. Mesenchymal stem cells will be obtained following the protocol proposed by Ouryazdanpanah et al [33] The advantages of this protocol are that it uses a small amount of human peripheral blood sample, avoid the necessity of mobilization with granulocyte-colony stimulating factor (G-CSF), and yields a large number of mesenchymal stem cells.…”

Section: Search Of Signature Of Epigenetic Alterations In Experimental Model and Clinical Cases Of Zikvassociated Microcephalymentioning

confidence: 99%

“…Cells regarded mesenchymal stem cells will be those expressing CD90, CD105, and CD73 and not expressing CD45 [33]. Similarly, hematopoietic stem cells will be selected using the method reported by Lemos et al [34] to quantify CD34+ cells. These CD34+ cells will be obtained from non-mobilized peripheral blood, which is sufficient to carry out the epigenomics analyses of this project [35].…”

Section: Search Of Signature Of Epigenetic Alterations In Experimental Model and Clinical Cases Of Zikvassociated Microcephalymentioning

confidence: 99%

Co-factor Influences the Severity of Zika Virus-Associated Microcephaly

Nogueira¹,

Andrade²,

Zanini³

et al. 2021

Preprint

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Microcephaly has been regarded the most remarkable consequence of the Zika virus (ZIKV) epidemic in Brazil 2015. It remains to be determined whether there are factors that contribute to the degree of brain lesion associated with ZIKV infection during pregnancy. Previous studies showed that socioeconomic conditions correlate with ZIKV-associated microcephaly. Certain nutritional deficits display the potential to interfere in the mechanistic target of rapamycin (mTOR) signaling, which plays a major role in the pathophysiology of ZIKV-associated microcephaly. We hypothesize that a nutritional or environmental co-factor that interferes in mTOR signaling correlates with ZIKV-associated birth defects. To assess this hypothesis, we plan to: 1) develop a mouse model of ZIKV-associated microcephaly through intravenous injection of ZIKV and rapamycin for a straightforward interference on mTOR receptor; 2) determine in the experimental model and in cases of ZIKV-associated microcephaly the epigenetic signature (DNA methylation pattern) in neurons and muscle cells harvested by biopsy, and in hematopoietic and mesenchymal stem cells sorted from blood; 3) analyze through mass spectrometry in serum of pregnant female mice submitted to ZIKV and rapamycin injection and in serum of mothers of children with ZIKV-associated microcephaly the metabolomic pattern of cholesterol (a nutritional status marker), vitamin A and its metabolite retinoic acid, folate, and other metabolites related to these three nutritional factors; 4) check whether pregnant female mice submitted to intravenous injection of ZIKV and feed with a deficient diet of the most likely co-factor found in this study give birth to microcephalic mice with features that mimic clinical cases. In summary, our general objective is to develop an experimental model that mimics ZIKV-associated microcephaly cases and to find a co-factor involved in the microcephaly outbreak in Brazil 2015.

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Quantification of peripheral blood CD34+ cells prior to stem cell harvesting by leukapheresis: a single center experience

Cited by 16 publications

References 24 publications

Pattern of autologous stem cell transplants at a tertiary care government hospital, with emphasis on transplant outcomes with pre-harvest CD34+ level

Pattern of autologous stem cell transplants at a tertiary care government hospital, with emphasis on transplant outcomes with pre-harvest CD34+ level

A novel small molecule CXCR4 antagonist potently mobilizes hematopoietic stem cells in mice and monkeys

Co-factor Influences the Severity of Zika Virus-Associated Microcephaly

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