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CD34 Enumeration, How and When 2011 - Webinar Recording (Nov 2011)

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CD34 Enumeration, How and When 2011

Organized by the ISCT Lab Practices Committee

Chair and Speaker: Varda Deutsch, PhD, Hematology Laboratory Director, Tel-Aviv Sourasky Medical Center

Speaker: Frank Preijers, PhD, Stem Cell Laboratory Director/Head section Immunophenotyping, Radboud University Nijmegen Medical Center


CD34+ cell enumeration to assess the quality of stem cell transplants is still a subject for improvement. Although the CV values in different external quality assessment schemes of CD34 are decreasing during the last decade, as a result of standardization and education as well as better reagents and flow cytometers, in many stem cell laboratories this technique still needs attention. Generally, the determination of the number of CD34+ cells in fresh cell sources can be performed reliably. Yet there are differences using different reagents. The results are often worse in samples after cryopreservation and in samples with less than optimal viability. Mainly in these determinations the enumeration assay needs improvement. Therefore the applied methods, the used CD34 antibodies and fluorochromes, the gating procedure (especially the sequential back gating on CD45), the right use of 7-AAD when and application of the counting beads may introduce deviations. Additionally, the flow cytometer used as well as the calibration of lasers and PMTs, and the compensation of spectral overlap between fluorochromes may influence strongly the outcome of the determinations. The most important parameters and issues that will influence the final CD34 count will be discussed.


  • Review methods and guidance documents for CD 34 enumeration in grafts.
  • Discuss reagents, gating, viability testing, fluorochromes and overlap between fluorochromes, calibration of lasers and problematic samples.
  • Emphasize the most important issues that influence the final CD34+ cell counts.


This product is a downloadable PDF containing a link to the mp4 video recording of this presentation featuring the Power Point slides and audio recording. 

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