title: Comparison of single- and dual-platform approaches to enumerate CD34(+) cells in bone marrow and mobilized peripheral blood stem cells.
abstract: BACKGROUND
  Different flow cytometric methods have been developed to derive absolute CD34(+) cells in predicting transplant outcome. Two techniques for preparing cells for quantification of CD34(+) cells were compared.
  
  
  METHODS
  Enumeration of CD34(+) cells in 16 samples of bone marrow (BM) and in 29 samples of mobilized peripheral blood stem cells (PBSC) obtained by leukapheresis was assessed simultaneously by single-platform (ProCOUNT kit) and dual-platform (Milan protocol) approaches within the first 3 h of collection.
  
  
  RESULTS
  Absolute number of CD34(+) cells obtained in PBSC and BM using single- and dual-platform methods showed high determination coefficients as follows: for PBSC, slope = 1.0515 +/- 0.048, y-intercept = 88.638 +/- 52.45, and r(2) = 0.941, and for BM, slope = 1.0203 +/- 0.093, y-intercept = 122.25 +/- 20.65, and r(2) = 0.878. There were no statistically significant differences in absolute number of CD34(+) cells from PBSC between single-platform (mean 575/microL, range 70-3683/microL) and dual-platform (786/microL, range 51-3804/microL) assays. In contrast, absolute number of CD34(+) cells from BM was significantly lower (p = 0.0002) when enumerated by ProCount kit (135/microL, 14-758/microL) than with dual-platform method (260/microL, 74-889/microL).
  
  
  CONCLUSIONS
  Both approaches can be used indistinctly to estimate absolute number of CD34(+) cells in PBSC but not in BM.
authors.affiliation: Departamento de Hematología y Oncología, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico. piedras@quetzal.innsz.mx
authors.forename: Josefa
authors.initials: J
authors.lastname: Piedras-Ross
authors.forename: Eucario
authors.initials: E
authors.lastname: León-Rodríguez
authors.forename: Sergio
authors.initials: S
authors.lastname: Sánchez-Guerrero
authors.forename: Xavier
authors.initials: X
authors.lastname: López-Karpovitch
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis*; D001854:Bone Marrow Cells / Q000166:cytology*; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D013223:Statistics as Topic; D013234:Stem Cells / Q000502:physiology*
chemical_list: D018952:Antigens, CD34