title: Simultaneous use of rhodamine 123, phycoerythrin, Texas red, and allophycocyanin for the isolation of human hematopoietic progenitor cells.
abstract: The supravital, mitochondrial specific dye Rhodamine 123 (R123) was used in conjunction with three monoclonal antibodies to isolate a population of human bone marrow (BM) cells enriched for hematopoietic progenitor cells. BM cells stained with phycoerythrin-HLA-DR, Texas red-CD34, allophycocyanin-CD15, and R123 were fractionated using four-color immunofluorescence cell sorting. Cells expressing CD34 but not HLA-DR and CD15 (CD34+ HLA-DR- CD15-) were subdivided according to their reactivity with R123 into quiescent, R123 dull (R+) or cycling, R123 bright (R++) subpopulations. Morphological analysis and hematopoietic progenitor cell assays indicated that CD34+ HLA-DR- CD15- R+ cells contained larger numbers of blast cells and colony forming units than CD34+ HLA-DR- CD15- R++ cells. The flow cytometer settings used to accommodate the detection of the R123 fluorescence in combination with that of three other fluorochromes are described.
authors.affiliation: Department of Medicine, Hematology/Oncology, Indiana University School of Medicine, Indianapolis 46202.
authors.forename: E F
authors.initials: EF
authors.lastname: Srour
authors.forename: T
authors.initials: T
authors.lastname: Leemhuis
authors.forename: J E
authors.initials: JE
authors.lastname: Brandt
authors.forename: K
authors.initials: K
authors.lastname: vanBesien
authors.forename: R
authors.initials: R
authors.lastname: Hoffman
mesh_terms: D000911:Antibodies, Monoclonal; D001854:Bone Marrow Cells; D002453:Cell Cycle; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006412:Hematopoietic Stem Cells; D006801:Humans; D010798:Phycocyanin; D010799:Phycoerythrin; D020112:Rhodamine 123; D012235:Rhodamines; D014966:Xanthenes
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D012235:Rhodamines; D014966:Xanthenes; C002497:allophycocyanin; D010798:Phycocyanin; D010799:Phycoerythrin; D020112:Rhodamine 123; C034657:Texas red