Improvement of the precision in CFU-GM and BFU-E counting by flow cytometry-based standardization of short-term culture assays.
The counting of colony-forming units granulocyte-macrophage (CFU-GM) and burst-forming units erythrocyte (BFU-E) provides substantial in vitro information about the graft quality after peripheral stem cell transplantation (PBSCT). By using different techniques for culturing and scoring, high inter- and intralaboratory coefficients of variation (CV) are frequently reported. We minimized the imprecision by using flow cytometry-based incorporation of constant numbers of CD34(+) cells per culture dish instead of the formerly used mononuclear cells. Our results show acceptable CVs for CFU-GM (12.3%) and for BFU-E (13.3%) based on this seeding technique, which contributes to fulfilling the demands of a quality assurance system in stem cell laboratories.
Sheikhzadeh
S
S
Institute of Immunology and Transfusion Medicine, University of Lübeck School of Medicine, Lübeck, Germany.
Hammers
H J
HJ
Hartwig
D
D
Kirchner
H
H
Schlenke
P
P
D018952:Antigens, CD34 / Q000032:analysis; D001772:Blood Cell Count / Q000295:instrumentation / Q000592:standards; D018929:Cell Culture Techniques; D015672:Erythroid Precursor Cells / Q000166:cytology*; D005434:Flow Cytometry / Q000592:standards*; D006098:Granulocytes / Q000166:cytology*; D006801:Humans; D008264:Macrophages / Q000166:cytology*; D008863:Microspheres; D023461:Myeloid Progenitor Cells / Q000166:cytology*; D015203:Reproducibility of Results
D018952:Antigens, CD34