Purification of human granulocyte-macrophage progenitor cells (CFU-C) using indirect immunofluorescence and cell sorting.
We have attempted to purify human bone marrow granulocyte-macrophage progenitor cells utilizing purified stimulating factor, indirect immunofluorescence and cell sorting. These cells can be labelled by sequential incubation with colony-stimulating factor (CSF), rabbit anti-CSF antibody and fluorescently labelled goat anti-rabbit gamma globulin. Flow photometry demonstrated that binding of CSF to marrow cells varied with CSF concentration and duration of incubation. Marrow cells were pre-concentrated by incubation with carbonyl iron followed by Percoll gradient centrifugation and then labelled by this indirect immunofluorescence method. Sorting of these cells yielded up to 95-fold purification of colony-forming cells and fractions containing in excess of 10% colony- and cluster-forming cells.
D001854:Bone Marrow Cells; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D003114:Colony-Forming Units Assay; D003115:Colony-Stimulating Factors / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006098:Granulocytes; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells; D006801:Humans; D008264:Macrophages
D003115:Colony-Stimulating Factors