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Fractionation of normal and beta-thalassemic human hemopoietic progenitor cells by immunomagnetic beads
1Cancer Research Unit, Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia.
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.
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.