Semi-automated flow cytometric analysis of CD34-expressing hematopoietic cells in peripheral blood progenitor cell apheresis products
Article first published online: 25 APR 2002
Volume 39, Issue 11, pages 1220–1226, November 1999
How to Cite
Gutensohn, K., Carrero, I., Krueger, W., Kroeger, N., Schäfer, P., Luedemann, K. and Kuehnl, P. (1999), Semi-automated flow cytometric analysis of CD34-expressing hematopoietic cells in peripheral blood progenitor cell apheresis products. Transfusion, 39: 1220–1226. doi: 10.1046/j.1537-2995.1999.39111220.x
- Issue published online: 25 APR 2002
- Article first published online: 25 APR 2002
- Received: 06 January 1998; Accepted: 17 March 1999
- BDIS = Becton Dickinson Immunocytometry Systems;
- FSC = forward scatter;
- FITC = fluorescein isothiocyanate;
- GRP = German reference protocol;
- ISHAGE = International Society of Hematotherapy and Graft Engineering;
- MoAb(s) = monoclonal antibody(ies);
- PBPC(s) = peripheral blood progenitor cell(s);
- PBS = phosphate-buffered saline;
- PE = phycoerythrin;
- RBC(s) = red cell(s);
- RT = room temperature;
- SSC = side scatter;
- WBC(s) = white cell(s)
BACKGROUND: The measurement of CD34+ cells is the most important step in the quality control of peripheral blood progenitor cell apheresis products. For this purpose, flow cytometry is applied. Recently, a new test kit has been introduced for the enumeration of CD34-expressing cells, in combination with software support for semi-automation of data acquisition and analysis.
STUDY DESIGN AND METHODS: This study evaluated the ProCOUNT kit. Ninety samples obtained from peripheral blood progenitor cell apheresis products from 39 patients with hemato-oncologic diseases were analyzed. For data acquisition and analysis, ProCOUNT software was used. Data comparison was performed with parallel measurements according to the International Society for Hematotherapy and Graft Engineering (ISHAGE) guidelines and the German reference protocol for analysis of CD34-expressing cells.
RESULTS: Correlation of the German and ISHAGE techniques was excellent (r2 = 0.99). The initial correlation coefficient of ProCOUNT analysis with the German protocol was r2 = 0.89. In 21 (23.3%) of 90 ProCOUNT analyses, a warning message was encountered from the ProCOUNT software. Following manual reevaluation of these data with CellQUEST software, a correlation of r2 = 0.96 with the German protocol and r2 = 0.97 with the ISHAGE analyses was obtained. ANOVA testing revealed significant differences between ProCOUNT and ISHAGE techniques (p<0.05) and between ProCOUNT and the German protocol (p<0.05). No statistically significant difference between ISHAGE and German protocol was observed (p = 0.19).
CONCLUSION: The ProCOUNT kit and software for semi-automated data acquisition and analysis represents a further step toward standardization of CD34 cell quantitation in peripheral blood progenitor cell apheresis products. However, the occurrence of software warnings is high, and analysis or data reevaluation by experienced staff is still mandatory. Therefore, currently there is no definite advantage of the kit and software over the existing guidelines for CD34+ analysis in peripheral blood progenitor cell grafts.