Comparison of Single- and Dual-Platform Approaches to Enumerate CD34+ Cells in Bone Marrow and Mobilized Peripheral Blood Stem Cells

Piedras-Ross, Josefa; Sánchez-Guerrero, Sergio Arturo; López‐Karpovitch, Xavier

doi:10.1016/s0188-4409(02)00451-4

Cited by 8 publications

(3 citation statements)

References 16 publications

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“…Moretti and coworkers 13 reported a good correlation between the ISHAGE protocol and a commercial single‐platform system (ProCOUNT, BD Biosciences, San Jose, CA) when analyzing peripheral blood and leukapheresis samples, as long as the white blood cell (WBC) count did not exceed 25 × 10 9 per L. Correlation was reported as suboptimal when analyzing cord blood samples. Piedras‐Ross and coworkers 10 also found a good correlation between the use of the dual‐platform Milan protocol 5 and the single‐platform ProCOUNT method in peripheral blood and apheresis products, but an underestimation of CD34+ cells was observed by the use of the single‐platform method in marrow samples 14 . In contrast, Keeney and colleagues 11 found that the single‐platform approaches might overestimate the numbers of CD34+ cells as a result of the “vanishing counting bead phenomenon” that is observed when the protein content of the sample 15 is low.…”

Section: Discussionmentioning

confidence: 93%

Instrument‐ and protocol‐dependent variation in the enumeration of CD34+ cells by flow cytometry

et al. 2006

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Section: Discussionmentioning

confidence: 93%

Instrument‐ and protocol‐dependent variation in the enumeration of CD34+ cells by flow cytometry

et al. 2006

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“…Identification of total MPs and their subsets is crucial to assess an accurate MP quantification, for the establishment of the MP role as markers of cardiovascular risk, disease and outcome in diabetic patients. The application of a common dual-platform approach (42), usually based on the simultaneous leucocyte count, is not recommended for MPs due to enormous differences in sizes between MPs and leukocytes. Single-platform methods are based on the use of commercial count beads, characterised by sizes substantially higher than that of the largest MPs, therefore requiring appropriate adjustment of the flow cytometry plot scales to correctly visualise count beads.…”

Section: Flow Cytometry Identification and Enumeration Of Microparticlesmentioning

confidence: 99%

Microparticles as new markers of cardiovascular risk in diabetes and beyond

Santilli¹,

Marchisio²,

Lanuti³

et al. 2016

Thromb Haemost

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SummaryThe term microparticle (MP) identifies a heterogeneous population of vesicles playing a relevant role in the pathogenesis of vascular diseases, cancer and metabolic diseases such as diabetes mellitus. MPs are released by virtually all cell types by shedding during cell growth, proliferation, activation, apoptosis or senescence processes. MPs, in particular platelet-and endothelial-derived MPs (PMPs and EMPs), are increased in a wide range of thrombotic disorders, with an interesting relationship between their levels and disease pathophysiology, activity or progression. EMP plasma levels have been associated with several cardiovascular diseases and risk factors. PMPs are also shown to be involved in the progressive formation of atherosclerotic plaque and development of arterial thrombosis, especially in diabetic patients. Indeed, diabetes is characterised by an increased procoagulant state and by a hyperreactive platelet phenotype, with enhanced adhesion, aggregation, and activation. Elevated MP levels, such as TF + MPs, have been shown to be one of the procoagulant determinants in patients with type 2 diabetes mellitus. Atherosclerotic plaque constitutes an opulent source of sequestered MPs, called "plaque" MPs. Otherwise, circulating MPs represent a TF reservoir, named "bloodborne" TF, challenging the dogma that TF is a constitutive protein expressed in minute amounts. "Blood-borne" TF is mainly harboured by PMPs, and it can be trapped within the developing thrombus. MP detection and enumeration by polychromatic flow cytometry (PFC) have opened interesting perspectives in clinical settings, particularly for the evaluation of MP numbers and phenotypes as independent marker of cardiovascular risk, disease and outcome in diabetic patients.

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“…2 Bland-Altman plot of percent difference comparing CD34+ hematopoietic stem cell count results obtained with both SP and DP approaches assays, such as intra-and inter-observer reproducibility and sampling errors [22]. Most of the published studies in this regard were based on hematological analyzers that operate on the Coulter principle to perform the DP technique, as Sysmex KX-21 for Ngoma A.M et al [23], a Coulter counter STKS model for Piedras-Ross J et al [24], and Sysmex k-1000, Technicon H3 for Gratama JW et al [4]. All these hematological analyzers, which operation is based on the impedance principle, were reported to have limitations against the presence of nRBC.…”

Section: Discussionmentioning

confidence: 99%

Enumeration of CD34+ haemopoietic stem cells: comparative study of the performance of the SYSMEX XN-1000 hematology analyzer in a dual-platform approach versus a single-platform approach

et al. 2021

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Data regarding the quantification of CD34+ hematopoietic stem cell HSC in dual-platform (DP) approach using a hematological analyzer featuring dedicated nRBC channel are limited if not inexistent. The objective of this study is to compare the performance of the DP approach on quantification of CD34+ HSC using a new generation of hematological analyzer featuring dedicated nRBC channel, Sysmex XN-1000, with the single-plate (SP) form approach using Beckman Coulter's Navios flow cytometer. Using the International Society for Hematotherapy and Transplantation protocol, a total of 86 samples were analyzed in multiple myeloma patients planned for autologous HSC transplantation. The Beckman Coulter's Stem-Kit™ reagents were used to perform both approaches DP and SP. The count of CD34+ cells of the SP deduced directly from the software that manage the Navios flow cytometry was compared to CD34+ cell count obtained by DP. The DP approach was made by combining flow cytometry analysis and the Sysmex XN-1000 hematological cell analyzer data. The analysis of the agreement of the results between these two approaches using the Bland-Altman diagram gave a bias of one cell/μl. As for the equation of the right of Passing-Bablok, it is Y (SP) = 0.166667 + 0.969697 X (DP). The slight differences in CD34+ stem cell counting between these two approaches did not achieve statistical significance. Both SP and DP approaches were effective and yield similar results. DP seems to be a simple and affordable approach that is well suited to the needs of laboratories where this type of analyzer is available.

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Comparison of Single- and Dual-Platform Approaches to Enumerate CD34+ Cells in Bone Marrow and Mobilized Peripheral Blood Stem Cells

Cited by 8 publications

References 16 publications

Instrument‐ and protocol‐dependent variation in the enumeration of CD34+ cells by flow cytometry

Instrument‐ and protocol‐dependent variation in the enumeration of CD34+ cells by flow cytometry

Microparticles as new markers of cardiovascular risk in diabetes and beyond

Enumeration of CD34+ haemopoietic stem cells: comparative study of the performance of the SYSMEX XN-1000 hematology analyzer in a dual-platform approach versus a single-platform approach

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