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True volumetric counting of CD34+ cells using flow cytometry.
Rico, Laura G; Bardina, Jorge; Salvia, Roser; Ward, Michael D; Bradford, Jolene A; Petriz, Jordi.
Afiliación
  • Rico LG; Functional Cytomics Lab, Germans Trias i Pujol Research Institute (IGTP), ICO-Hospital Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona (Barcelona), Spain.
  • Bardina J; Functional Cytomics Lab, Germans Trias i Pujol Research Institute (IGTP), ICO-Hospital Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona (Barcelona), Spain.
  • Salvia R; Functional Cytomics Lab, Germans Trias i Pujol Research Institute (IGTP), ICO-Hospital Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona (Barcelona), Spain.
  • Ward MD; Thermo Fisher Scientific, Fort Collins, CO, USA.
  • Bradford JA; Thermo Fisher Scientific, Fort Collins, CO, USA.
  • Petriz J; Functional Cytomics Lab, Germans Trias i Pujol Research Institute (IGTP), ICO-Hospital Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona (Barcelona), Spain. Electronic address: jpetriz@igtp.cat.
J Immunol Methods ; 527: 113649, 2024 04.
Article en En | MEDLINE | ID: mdl-38395105
ABSTRACT
While the single-platform flow cytometric CD34+ cell counting method is the preferred choice to predict the yield of mobilized peripheral blood stem cells, most flow cytometers lack the ability of hematology counter analyzers to perform volumetric counting. However, one of the problems using reference microbeads is the vanishing counting bead phenomenon. This phenomenon results in a drop in microbeads concentration and reduces the total and relative number of beads in calibration procedures. In the last years, flow cytometers including a volumetric system to quantify cells have been developed and may represent a promising alternative to enumerate CD34+ cells avoiding the use of beads. In this study we have used a direct true volumetric counting of CD34+ cells under continuous flow pump to overcome potential drawbacks with impact in rare cell analysis. To confirm this hypothesis, we have compared the results of CD34+ cell enumeration using non-volumetric vs. volumetric systems with FC500 (Beckman Coulter) and Attune NxT (ThermoFisher) flow cytometers, respectively, in mobilized peripheral blood samples. No statistically significant differences were observed between measurements of CD34+ cells using beads, when the FC500 and Attune NxT absolute counting values were compared, or when CD34+ counts were compared on the Attune NxT, either using or not using beads. Linear regressions to study the relationship between volumetric and non-volumetric CD34+ counts confirmed the accuracy of each method. Bland-Altman test showed agreement between both methods. Our data showed that CD34+ cell enumeration using a volumetric system is comparable with current counting systems. This method represents an alternative with the advantage of the simplification of sample preparation and the reduction of the analysis subjectivity.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Citometría de Flujo Idioma: En Revista: J Immunol Methods Año: 2024 Tipo del documento: Article País de afiliación: España Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Citometría de Flujo Idioma: En Revista: J Immunol Methods Año: 2024 Tipo del documento: Article País de afiliación: España Pais de publicación: Países Bajos