Cell cycle-dependent reactivity with the monoclonal antibody Ki-67 during myeloid cell differentiation.
The specificity and sensitivity of the monoclonal antibody Ki-67 in identifying proliferating cell compartments was tested with the human promyelocytic leukemia cell line HL-60 using multi-parameter flow cytometry. While correlated measurements of DNA content and Ki-67 immunofluorescence indicated that the antigen was present in all phases of the cell cycle, reactivity with the antibody was highest in proliferating S and G2+M cells. The analysis of the BrdU content of cells sorted on the basis of reactivity with Ki-67 showed a correlation between Ki-67 reactivity and BrdU uptake. In HL-60 cells induced to differentiate with dimethyl sulfoxide (DMSO), the loss of reactivity with Ki-67 paralleled the exit of cells from the cell cycle. This was not observed in DMSO-resistant HL-60 cells. These results validate the usefulness of the Ki-67 antibody for determining the proliferative stage of mammalian cells in culture.
D000911:Antibodies, Monoclonal; D001973:Bromodeoxyuridine / Q000378:metabolism; D002451:Cell Compartmentation; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D004273:DNA, Neoplasm / Q000032:analysis; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology*
D000911:Antibodies, Monoclonal; D004273:DNA, Neoplasm; D001973:Bromodeoxyuridine; D004121:Dimethyl Sulfoxide