title: Performance of a novel BD stem cell enumeration kit on two flow cytometry systems.
abstract: BACKGROUND
  Hematopoietic stem cell transplantation, which requires accurate enumeration of stem cells, is routinely used in clinical settings. Flow cytometry provides a qualitative and quantitative assessment of CD34⁺ cells. Precision, linearity, and stability of the novel BD™ Stem Cell Enumeration (SCE) Kit were evaluated on two flow cytometry platforms using a modified ISHAGE gating strategy and including a viability dye for data acquisition and analysis.
  
  
  METHODS
  Precision and linearity were evaluated on BD FACSCanto™ II and BD FACSCalibur™ systems. Stability was evaluated on the BD FACSCanto II system. Precision was tested using both high and low controls. Linearity was evaluated using dilutions from CD34⁺ cell pools, while stability was evaluated using fresh leukapheresis specimens.
  
  
  RESULTS
  Both systems showed precision with limited variability in absolute counts and percentages of viable CD34⁺ cells. The linearity range of viable CD34⁺ cells in both systems was established at 0-1000 cells/μL, showing a linear relationship (R² = 0.99). Stability of CD34⁺ cells in mobilized leukapheresis samples was confirmed up to 24 h after collection and up to 60 min after the end of stain/lyse procedures.
  
  
  CONCLUSIONS
  The BD SCE Kit on both flow cytometry systems shows consistent and reproducible results.
authors.affiliation: BD Biosciences, San Jose, CA 95131, USA. Imelda_Omana-Zapata@bd.com
authors.forename: I
authors.initials: I
authors.lastname: Omana-Zapata
authors.forename: F
authors.initials: F
authors.lastname: Oreizy
authors.forename: F
authors.initials: F
authors.lastname: Mosqueda
authors.forename: M
authors.initials: M
authors.lastname: Salemenik
authors.forename: A
authors.initials: A
authors.lastname: Lin
authors.forename: J L
authors.initials: JL
authors.lastname: Wen
authors.forename: R
authors.initials: R
authors.lastname: Varro
authors.forename: L
authors.initials: L
authors.lastname: Lee-Haynes
authors.forename: O G
authors.initials: OG
authors.lastname: Segurado
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002138:Calibration; D002452:Cell Count / Q000592:standards; D002470:Cell Survival; D005434:Flow Cytometry / Q000592:standards*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007937:Leukapheresis; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D015203:Reproducibility of Results
chemical_list: D018952:Antigens, CD34; D015415:Biomarkers; D011933:Reagent Kits, Diagnostic