Semi-automated flow cytometric analysis of CD34-expressing hematopoietic cells in peripheral blood progenitor cell apheresis products

• Published in: Transfusion (Philadelphia, Pa.) Pa.), 1999-11, Vol.39 (11), p.1220-1226
• Main Authors: Gutensohn, K., Carrero, I., Krueger, W., Kroeger, N., Schäfer, P., Luedemann, K., Kuehnl, P.
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Antigens, CD34 - blood; BDIS = Becton Dickinson Immunocytometry Systems; Biological and medical sciences; Blood Component Removal; Bone marrow, stem cells transplantation. Graft versus host reaction; Female; FITC = fluorescein isothiocyanate; Flow Cytometry - methods; FSC = forward scatter; GRP = German reference protocol; Hematopoietic Stem Cell Mobilization; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - immunology; Humans; Image Processing, Computer-Assisted; ISHAGE = International Society of Hematotherapy and Graft Engineering; Male; Medical sciences; Middle Aged; 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; Software; SSC = side scatter; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; WBC(s) = white cell(s)
• ISSN: 0041-1132; 1537-2995
• DOI: 10.1046/j.1537-2995.1999.39111220.x

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