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 Citation (Scopus)

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

Fingerprint

Leukapheresis
blood cells
stem cells
blood
assays
cells
Peripheral Blood Stem Cell Transplantation
bone marrow transplant
cell transplantation
Blood Component Removal
Autologous Transplantation
Fluorescein-5-isothiocyanate
Hematologic Neoplasms
Bone Marrow Transplantation
methodology
standardization
Quality Control
cell viability
quality control
flow cytometry

Keywords

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

ASJC Scopus subject areas

  • Hematology
  • Immunology
  • Food Science

Cite this

Precise CD34+ quantification using a multi-parameter flow-cytometric method with fluorescent microparticles. / Schlenke, P.; Frohn, C.; Müller-Steinhardt, M.; Hammers, H. J.; Saballus, M.; Klüter, H.

In: Infusionstherapie und Transfusionsmedizin, Vol. 26, No. 3, 01.05.1999, p. 172-178.

Research output: Contribution to journalArticle

Schlenke, P, Frohn, C, Müller-Steinhardt, M, Hammers, HJ, Saballus, M & Klüter, H 1999, 'Precise CD34+ quantification using a multi-parameter flow-cytometric method with fluorescent microparticles', Infusionstherapie und Transfusionsmedizin, vol. 26, no. 3, pp. 172-178.
Schlenke, P. ; Frohn, C. ; Müller-Steinhardt, M. ; Hammers, H. J. ; Saballus, M. ; Klüter, H. / Precise CD34+ quantification using a multi-parameter flow-cytometric method with fluorescent microparticles. In: Infusionstherapie und Transfusionsmedizin. 1999 ; Vol. 26, No. 3. pp. 172-178.
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