Simultaneous staining of exponentially growing versus plateau phase cells with the proliferation-associated antibody Ki-67 and propidium iodide: analysis by flow cytometry.
The antibody Ki-67, which detects proliferating cells, was used in combination with propidium iodide, a DNA-specific dye. The double-staining method allowed discrimination of cells in the phases of the cell cycle as well as the recognition of Ki-67 staining characteristics. Suspension cultures of U937 cells were measured in exponential growth and plateau phase in nutritional deprivation. The fraction of Ki-67 positive cells was nearly 100% 2 days after dilution and 46% 7 days after dilution of the cultures. Stathmokinetic measurements with colchicine and flow cytometry measurements with the BrdU-Hoechst technique yielded close to 100% proliferation at day 2 but only 18% and 6%, respectively, at day 7. The discrepancy between Ki-67 results and the results of the two other methods is considered to be a characteristic of nutritionally deprived cells.
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D002455:Cell Division; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology; D005434:Flow Cytometry; D007399:Interphase; D007700:Kinetics; D009687:Nuclear Proteins / Q000032:analysis / Q000276:immunology*; D010617:Phenanthridines; D011419:Propidium; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D009687:Nuclear Proteins; D010617:Phenanthridines; D011419:Propidium; D003078:Colchicine