Rapid detection of S-phase cells by anti-bromodeoxyuridine monoclonal antibody in 9L brain tumor cells in vitro and in situ.
FITC-conjugated anti-bromodeoxyuridine (BrdUrd) monoclonal antibody (anti-BU-MAb) was used to detect S-phase cells of 9L rat brain tumor cells in vitro and in situ. Monolayer 9L cells were treated with 0.625-20 microM of BrdUrd for 30 min, harvested, and reacted with a 1:100 dilution of FITC-conjugated anti-BU-MAb and analyzed with a flow cytometer. BrdUrd-treated cells stained satisfactorily with antibody. Values obtained for the labeling index using this method (48.6%) were 10%-20% higher than the fraction of cells in S-phase calculated from DNA histograms or as the labeling index calculated from autoradiographs of cells pulse-labeled with 3H-thymidine. BrdUrd (1-40 mg/kg) was administered by i.p. injection to rats bearing 9L brain tumors. Single cell suspensions obtained by disaggregation of excised tumors were stained with anti-BU-MAb. The percentage of fluorescent cells (15.9%) calculated using this method was similar to that of S-phase cells (17.2%) calculated from DNA histograms and from autoradiographics for tumor bearing rats pulse-labeled with 3H-thymidine in situ. The antibody staining technique is a rapid and accurate method for various cell kinetic studies both in vitro and in vivo in a rat model, and has promise as a technique for the study of cell kinetics in humans.
D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000473:pathology; D001932:Brain Neoplasms / Q000473:pathology*; D001973:Bromodeoxyuridine / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D004273:DNA, Neoplasm / Q000378:metabolism; D005434:Flow Cytometry; D007399:Interphase; D007700:Kinetics; D008856:Microscopy, Fluorescence; D009368:Neoplasm Transplantation; D051381:Rats; D011916:Rats, Inbred F344
D000911:Antibodies, Monoclonal; D004273:DNA, Neoplasm; D001973:Bromodeoxyuridine