Multiparameter flow-cytometrical quantitation of circulating CD34(+)-cells: correlation to the quantitation of circulating haemopoietic progenitor cells by in vitro colony-assay.
A multiparameter flow-cytometrical method for the quantitation of CD34(+)-cells present in adult human peripheral blood cells (PBMC) has been developed. PBMC from 13 healthy adult subjects were analysed for CD34(+)-cells by flow-cytometry, with only the lymphoid population of cells negative for anti-CD3-moAb included in the analysis. At the same time mononuclear cells from the same individuals were depleted of CD3(+)- and CD14(+)-cells by immunomagnetic purging. These cells were cultured for haemopoietic colonies. A correlation coefficient of 0.92 was calculated by regression analysis of CD34(+)-cells number versus the numbers of colonies (CFU-GM, BFU-E). Purging with anti-CD34-moAb abrogated colony-growth. The flow-cytometrical method for the determination of circulating CD34(+)-cells was used for the determination of circulating haemopoietic progenitor cells in one patient, in order to time leukapheresis for subsequent autologous reinfusion following high-dose chemotherapy. Determination of CD34(+)-cells from human adult peripheral blood by multiparameter flow-cytometry promises to be a useful tool for monitoring circulating haemopoietic progenitor cells as identified by anti-CD34-moAb.
Serke
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Hämatologische Ambulanz, Universitätsklinikum R. Virchow-Charlottenburg, Berlin, Germany.
Säuberlich
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Huhn
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D000328:Adult; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis*; D018952:Antigens, CD34; D000943:Antigens, Differentiation / Q000032:analysis*; D015214:Antigens, Differentiation, Myelomonocytic / Q000276:immunology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology; D001772:Blood Cell Count; D017252:CD3 Complex; D002469:Cell Separation / Q000379:methods; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry / Q000379:methods; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000276:immunology*; D006801:Humans; D018950:Lipopolysaccharide Receptors; D008297:Male; D011948:Receptors, Antigen, T-Cell / Q000276:immunology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018952:Antigens, CD34; D000943:Antigens, Differentiation; D015214:Antigens, Differentiation, Myelomonocytic; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D018950:Lipopolysaccharide Receptors; D011948:Receptors, Antigen, T-Cell