A flow cytometric method for measuring lymphocyte proliferation directly from tissue culture plates using Ki-67 and propidium iodide.
A new method for measuring lymphocyte proliferation in response to mitogens and allogeneic cells without using radiolabelling is described. It utilizes flow cytometry and the monoclonal antibody, Ki-67, which detects a nuclear proliferation antigen. The entire test is performed in standard, 96-well tissue culture plates. Stable, clean nuclear suspensions rather than whole cells were used to avoid non-specific staining. The nuclei were stained by the indirect fluorescent method. Simultaneous measurements of DNA content were possible by dual staining with propidium iodide (PI). The percentage of Ki-67-positive nuclei correlated well with [3H]thymidine uptake and morphologic quantitation of blasts. This method avoids use of radioactive material and is less time consuming.
D000911:Antibodies, Monoclonal; D002453:Cell Cycle; D002467:Cell Nucleus / Q000276:immunology; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D010835:Phytohemagglutinins / Q000494:pharmacology; D011043:Pokeweed Mitogens / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D010835:Phytohemagglutinins; D011043:Pokeweed Mitogens; D003208:Concanavalin A