Quality assessment of CD34+ stem cell enumeration: experience of the United Kingdom National External Quality Assessment Scheme (UK NEQAS) using a unique stable whole blood preparation


Dr Barnett UK NEQAS for Leucocyte Immunophenotyping, Department of Haematology, Royal Hallamshire Hospital, Glossop Road, Sheffield S10 2JF.


CD34+ peripheral blood stem cell (PBSC) mobilization and harvesting has rapidly replaced autologous bone marrow as a source of stem cells for transplantation. Timing and adequacy of harvests rely upon the accurate enumeration of circulating CD34+ cells. However, previous EQA programmes have reported interlaboratory CVs as high as 284%, suggesting the need for greater standardization. In addition the routine use of fresh and/or frozen cells as analytes also introduces antigen instability as a variable factor. To circumvent this problem and achieve a true reflection of interlaboratory variation, we have used a novel whole blood preparation in which the antigenic profiles of PBSCs, as determined by flow cytometry, are retained for > 200 d. This international scheme, currently the largest in the world, distributes aliquots of stabilized whole blood bi-monthly to 91 laboratories in 20 countries (44 U.K., 47 overseas). Participants are required to determine the percentage and absolute values for CD34+ PBSCs using in-house techniques. Adopting such a preparation, a more accurate determination of interlaboratory variation has been possible when compared to previous EQA studies, with CVs as low as 22% and 24% for percentage and absolute counts. In addition the programme has established that a wide range of methods are in routine use, emphasizing the urgent requirement for national/international consensus guidelines.