Analyses of sister-chromatid exchange and cell-cycle kinetics in mouse T- and B-lymphocytes from peripheral blood cultures.
A reliable mouse peripheral blood lymphocyte culture assay has been developed for sister-chromatid exchange analysis. Crucial aspects for optimal mitogenesis include: (1) the addition of 5 X 10(5) leucocytes/ml culture; (2) the use of animals with leucocyte counts from 5 to 7 X 10(6)/ml; and (3) the addition of 6% mouse plasma for the first 24 h of a total 54-h incubation. When 7 micrograms phytohemagglutinin/ml were used to stimulate T-lymphocytes, the mitotic index was 3.4 +/- 0.3%, 28 +/- 2.3% of the metaphases were in first-division, and the SCE frequency/metaphase was 7.3 +/- 0.2 (n = 14 mice). When B-lymphocytes were stimulated with 60 micrograms lipopolysaccharide/ml, the mitotic index was 4.5 +/- 0.3%, 64 +/- 3.3% of the metaphases were in first-division, and the SCE frequency/metaphase was 4.6 +/- 0.1 (n = 7 mice). This culture method consistently yields sufficient numbers of metaphases from both B- and T-lymphocytes for SCE and chromosome-breakage studies.
D000818:Animals; D001402:B-Lymphocytes / Q000648:ultrastructure*; D002453:Cell Cycle; D002478:Cells, Cultured; D003434:Crossing Over, Genetic; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D051379:Mice; D010835:Phytohemagglutinins / Q000494:pharmacology; D012854:Sister Chromatid Exchange; D013601:T-Lymphocytes / Q000648:ultrastructure*
D008070:Lipopolysaccharides; D010835:Phytohemagglutinins