Precise CD34+ quantification using a multi-parameter flow-cytometric method with fluorescent microparticles

P. Schlenke, C. Frohn, M. Müller-Steinhardt, H. J. Hammers, M. Saballus, H. Klüter

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Background: In recent years, peripheral blood stem cell transplantation has replaced autologous bone marrow transplantation to a large extent in patients with hematological malignancies. Optimal timing and adequate harvesting rely upon the precise quantification of mobilized CD34+ cells in blood and apheresis products. The quality assessment of collected stem cells includes viability tests, short-term culture assays and the determination of CD34+ cells by flow cytometry. External quality controls have suggested the urgent need for greater standardization of the enumeration of CD34+ cells. Material and Methods: We evaluated a single-platform assay for CD34+ cell quantification in peripheral blood and leukapheresis samples (n = 22) by using a four-parameter flow-cytometric method (forward and side angle light scatter, CD34 PE, CD45 FITC) including Flow Count fluorospheres for direct quantification. Results: When processing times of 30 and 300 s were compared, the interassay coefficients of variation declined significantly from 11% and 12% in peripheral blood and leukapheresis to 4.7% and 5.2% (p = 0.003 and p = 0.016), respectively. The precision analysis of the flow-cytometric method (intra-assay variation) generated coefficients of variation, which dropped from 8.1% and 8.5%, respectively, to 3.3% for both samples with regard to the processing time (p < 0.001). Conclusion: The data demonstrate the need of single-platform flow-cytometric methods based on fluorescent microparticles. They showed a greater precision of CD34+ cell enumeration over the course of analysis time. Because of its clinical relevance in peripheral blood stem transplantation, these improvements should supplement the current guidelines.

Original languageEnglish (US)
Pages (from-to)172-178
Number of pages7
JournalInfusionstherapie und Transfusionsmedizin
Volume26
Issue number3
StatePublished - May 1 1999

Keywords

  • CD34
  • Flow count
  • Flow cytometry
  • Microparticles
  • Quantification

ASJC Scopus subject areas

  • Food Science
  • Immunology
  • Hematology

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