Performance of a novel BD Stem Cell Enumeration Kit on two flow cytometry systems

• Published in: International journal of laboratory hematology Vol. 35; no. 4; pp. 393 - 399
• Main Authors: Omana-Zapata, I., Oreizy, F., Mosqueda, F., Salemenik, M., Lin, A., Wen, J. L., Varro, R., Lee-Haynes, L., Segurado, O. G.
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.08.2013
• Subjects: Antigens, CD34 - metabolism; Biomarkers - metabolism; Calibration; Cell Count - standards; Cell Survival; Engraftment; flow cytometry; Flow Cytometry - standards; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Humans; Leukapheresis; progenitor cells; Reagent Kits, Diagnostic - standards; Reproducibility of Results; stem cells; transplantation; Engraftment; stem cells; progenitor cells; flow cytometry; transplantation
• ISSN: 1751-5521; 1751-553X
• DOI: 10.1111/ijlh.12029

Summary Introduction 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. Conclusion The BD SCE Kit on both flow cytometry systems shows consistent and reproducible results.