Optimal conditions in [3H]-thymidine uptake studies to prevent radiation damage to cells. A scintimetric and cytofluorographic analysis.
Cells subjected to nucleoside incorporation studies using radiolabelled materials may suffer radiation damage that can alter the results. We did scintimetric and cytofluorographic analysis to confirm this and to determine the optimal experimental doses of, and exposure times to, [3H]-TdR, in order to prevent or minimize such radiation damage to cells. We found that cultures should be pulsed with 0.125 microCi for 14 hr when stimulated with phytohaemagglutinin 0.125 microCi for 18 hr when stimulated with pokeweed mitogen, 0.5 microCi for 8 hr when stimulated with concanavalin A and 0.5 microCi for 8 hr when subject to allogeneic stimulus, in order to achieve optimal incorporation with minimal disturbance of the cell cycle.
D002453:Cell Cycle / Q000528:radiation effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004307:Dose-Response Relationship, Radiation; D006801:Humans; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000378:metabolism / Q000528:radiation effects*; D010779:Photofluorography; D013936:Thymidine / Q000378:metabolism*; D013997:Time Factors; D014316:Tritium
D014316:Tritium; D004247:DNA; D013936:Thymidine