Related Experiment Video
Updated: May 15, 2026
Simultaneous Assessment of Kinship, Division Number, and Phenotype via Flow Cytometry for Hematopoietic Stem and Progenitor Cells
Published on: March 24, 2023
A new approach to CD34+ hematopoietic progenitor cell counting
Pedro Marín1, Lluís Jover, Jordi Petriz
1Servei d'Hemoteràpia i Hemostàsia, Hospital Clínic, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain.
Background:
In the absence of a gold standard for hematopoietic progenitor counting, the intra-laboratory variation between commonly used strategies for progenitor assessment was compared.
Methods And Materials:
We used a pool of FITC-conjugated monoclonal antibodies (CD14, CD11b) and PE-CD34 to facilitate CD34 counting, excluding CD14(+)/CD11b(+) bright cells. We compared this protocol with other common methodologies, such as the single-staining approach, known as the Milan method, and the ISHAGE multiparameter method.
Results:
We show that the CD14/CD11b protocol is a valid approach to progenitor cell counting. Though different methods give different results for progenitor cell counting, Lin's coefficient shows high concordance among flow cytometry counts but a substantial difference with colony- forming unit counts. Both the ISHAGE and CD14/CD11b protocol give higher counts than the Milan method. Moreover, on revising the ISHAGE analysis, we described a rare population of cells with the CD34(+)CD45(neg) phenotype, which could have an impact in CD34 counting.
Conclusions:
We have designed a valid alternative approach for hematopoietic progenitor cell counting, and we show that different methods give different results.
