The kinetics of the formation of a G2 block from tritiated thymidine in phytohemagglutinin-stimulated human lymphocytes

Alan Pollack, C. B. Bagwell, G. L. Irvin, J. A. Jensen

Research output: Chapter in Book/Report/Conference proceedingChapter

15 Scopus citations

Abstract

Flow cytometry (FCM) was used to monitor the radiation effects promoted by incorporated tritiated thymidine (3H-TdR) on phytohemagglutinin (PHA)-stimulated human peripheral blood lymphocytes stained with propidium iodide (PI). Lymphocyte microcultures were continuously labeled or pulse-labeled for various periods of time with different 3H-TdR concentrations. Two types of DNA histogram analyses were performed on unperturbed and 3H-TdR perturbed lymphocytes. The data analyses consisted of statistical analyses between averaged groups of histograms (nonparametric analysis) and cell cycle analyses (parametric analysis) to determine the percentages of cells in GO + G1,S and G2 + M. The results showed that 3H-TdR when added to proliferating lymphocytes under certain conditions (both short-term continuous and pulse-labeling) caused a highly significant increase in the proportion of tetraploid (4C) cells by FCM, the increase in the proportion of 4C cells represented a block in G2 and the relative increase in the percentage of 4C cells was proportional to 3H-TdR incorporation which was proportional to labeling time and concentration. Therefore, it was concluded that short labeling times be used to minimize adverse radiation effects when 3H-TdR is used to assay substances affecting lymphocyte proliferation or in the estimation of cell cycle time.

Original languageEnglish
Title of host publicationCytometry
Pages57-64
Number of pages8
Volume1
Edition1
StatePublished - Dec 1 1980

ASJC Scopus subject areas

  • Medicine(all)

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