Fractionation of normal and beta-thalassemic human hemopoietic progenitor cells by immunomagnetic beads.
Depletion of adherent cells, followed by simultaneous immunomagnetic bead depletion of Leu 4+, Leu 7+, Leu 11+, Leu M1+, Leu M3+, B1+, WEM-G11+, and Glycophorin A+ cells from normal bone marrow mononuclear cells, consistently led to recoveries of erythroid and nonerythroid colony-forming cells of greater than 100% and enrichment of 13- to 99-fold. Similarly, the recovery of mixed erythroid colony-forming cells was greater than 100%, with enrichments of 7.5- to 262-fold. This simple procedure, when used on normal bone marrow and beta-thalassemic peripheral blood mononuclear cells, as well as providing significant enrichment, suggests that colony assays on unfractionated human mononuclear cells specifically underestimate the number of BFU-E, Mix-CFC, and nonerythroid-CFC present.
Kannourakis
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Cancer Research Unit, Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia.
Bol
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D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D001853:Bone Marrow / Q000473:pathology; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D004912:Erythrocytes / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D008280:Magnetics; D013789:Thalassemia / Q000097:blood*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface