CD34+ cell enumeration in peripheral blood and apheresis samples, using two laboratory diagnostic kits or an institutional protocol

Olivero, S; Alario, Thierry; Ladaique, Patrick; Haccoun, Martine; Viens, Patrice; Blaise, Didier; Chabannon, Christian

doi:10.1038/sj.bmt.1701583

Cited by 23 publications

(14 citation statements)

References 19 publications

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“…The use of the BDIS ProCount kit was shown to reduce remarkably both the intralaboratory and interlaboratory assay variability (60,178,179). The same was demonstrated for the Beckman-Coulter StemKit assay in a singlecenter study (179).…”

Section: Multicenter Studies On Cd34؉ Cell Counting: the Emerging Conmentioning

confidence: 75%

Cytofluorometric methods for assessing absolute numbers of cell subsets in blood

et al. 2000

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The enumeration of absolute levels of cells and their subsets in clinical samples is of primary importance in human immunodeficiency virus (HIV)؉ individuals (CD4؉ T-lymphocyte enumeration), in patients who are candidates for autotransplantation (CD34؉ hematopoietic progenitor cells), and in evaluating leukoreduced blood products (residual white blood cells). These measurements share a number of technical options, namely, single-or multiple-color cell staining and logical gating strategies. These can be accomplished using single-or dual-platform counting technologies employing cytometric methods. Dual-platform counting technologies couple the percentage of positive cell subsets obtained by cytometry and the absolute cell count obtained by automated hematology analyzers to derive the absolute value of such subsets. Despite having many conceptual and technical limitations, this approach is traditionally considered as the reference method for absolute cell count enumeration. As a result, the development of single-platform technologies has recently attracted attention with several different technical approaches now being readily available. These single-platform approaches have less sources of variability. A number of reports clearly demonstrate that they provide better coefficients of variation (CVs) in multicenter studies and a lower chance to generate aberrant results. These methods are therefore candidates for the new gold standard for absolute cell assessments. The currently available technical options are discussed in this review together with the results of some cross-comparative studies. Each analytical system has its own specific requirements as far as the dispensing precision steps are concerned. The importance of precision reverse pipetting is emphasized. Issues still under development include the establishment of the critical error ranges, which are different in each test setting, and the applicability of simplified low-cost techniques to be used in countries with limited resources. Cytometry (Comm. Clin. Cytometry) 42:327-346, 2000.

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Section: Multicenter Studies On Cd34؉ Cell Counting: the Emerging Conmentioning

confidence: 75%

Cytofluorometric methods for assessing absolute numbers of cell subsets in blood

et al. 2000

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“…This commercially available diagnostic kit includes the viability dye 7-amino actinomycin D (7-AAD), that allows for a quantitation of viable CD34 þ and CD45 þ cells. 22 Absolute lymphocyte counts were determined using the Multitest kit (Becton Dickinson Immunocytometry Systems, San Jose, CA, USA). This commercially available diagnostic kit allows for a quantitation of 23 Data acquisitions and analyses were performed on a FACSCalibur four-color flow cytometer using the Cellquest and Multiset softwares (Becton Dickinson Immunocytometry System, BDIS, San Jose, CA, USA).…”

Section: Thawing Proceduresmentioning

confidence: 99%

CD34+ progenitors are reproducibly recovered in thawed umbilical grafts, and positively influence haematopoietic reconstitution after transplantation

Lemarié

Esterni

Calmels

et al. 2007

Bone Marrow Transplant

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Cord blood (CB) units are increasingly used for allogeneic transplantation. Cell dose, a major factor for CB selection, is evaluated before freezing by each CB bank, using various techniques. This may introduce variability and affect the prediction of cell recovery after thawing, or haematopoietic reconstitution. Forty-two children were transplanted at the same institution with unrelated CB units. All units were thawed and evaluated at the same cell therapy facility, using standard procedures. We investigated: (i) factors that affect cell loss after thawing, and (ii) the importance of CD34 þ cell doses. Prefreeze and postthaw CD34 þ cell doses were statistically correlated, thus suggesting that variability in numeration techniques used by different CB banks does not compromise the biological and clinical value of these figures. CD34 þ cell recovery appeared to be correlated with the absolute number of CD34 þ cells per frozen bag. Infused CD34 þ is the cell dose that better correlates with platelet reconstitution delay; in addition, when using a quartile comparison, haematopoietic recovery appeared to be related with prefreeze and post-thaw CD34 þ cell doses. We conclude that enumeration of CD34 þ cells in CB units is of biological significance, and may help select CB units and identify patients at risk of delayed recovery.

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“…Absolute counts of CD34 + cells were determined, using the Stem-Kit numeration kit (Beckman-Coulter/Immunotech, Hialeah, FL, USA), 23 according to the manufacturer's instructions. This commercially available diagnostic kit includes the viability dye 7-amino actinomycin D (7-AAD) that allows for a quantitation of both viable and nonviable CD34 + cells.…”

Section: Flow Cytometry Analysesmentioning

confidence: 99%

Preclinical evaluation of an automated closed fluid management device: CytomateTM, for washing out DMSO from hematopoietic stem cell grafts after thawing

Calmels

Houzé

et al. 2003

Bone Marrow Transplant

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Summary:Infusion of dimethylsulfoxide (DMSO) contained in cryopreserved and thawed hematopoietic stem cell (HSC) grafts is frequently associated with mild or moderate adverse reactions, and occasionally with more severe events including neurological symptoms. The severity of these complications is related to the amount of residual DMSO. We evaluated a recently available, closed, automated and 'cgmp (current good manufacturing practice) compliant' device (CytoMate TM ) for its ability to wash out DMSO at the expense of a limited loss of viable CD34 + cells. A total of 16 procedures were carried out with 39 blood HSC bags intended for destruction. Mean amounts of DMSO for each cellular product (one, two or three bags) were between 12.2 and 39.6 g before thawing; after the washing procedure, residual DMSO quantities were between 0.1 and 3.7 g. When set up to reproducibly allow for a more than 96% elimination of DMSO, processing of thawed cells with the CytoMate TM cell processor resulted in a mean recovery of viable total cells, CD34 + cells and lymphocyte subsets above 60%. We conclude that this simple and efficient washing technique is suitable for routine processing of HSC grafts. Clinical studies will demonstrate whether a reduction in the incidence of adverse effects associated with DMSO infusion is observed.

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CD34+ cell enumeration in peripheral blood and apheresis samples, using two laboratory diagnostic kits or an institutional protocol

Cited by 23 publications

References 19 publications

Cytofluorometric methods for assessing absolute numbers of cell subsets in blood

Cytofluorometric methods for assessing absolute numbers of cell subsets in blood

CD34+ progenitors are reproducibly recovered in thawed umbilical grafts, and positively influence haematopoietic reconstitution after transplantation

Preclinical evaluation of an automated closed fluid management device: CytomateTM, for washing out DMSO from hematopoietic stem cell grafts after thawing

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