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

Journal Article · Tue Jul 01 04:00:00 EDT 1980 · Cytometry (Baltimore); (United States)

DOI:https://doi.org/10.1002/cyto.990010112· OSTI ID:5440615

Pollack, A; Bagwell, C B; Irvin, G L; Jensen, J A

Flow cytometry (FCM) was used to monitor the radiation effects promoted by incorporated tritiated thymidine (/sup 3/H-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 /sup 3/H-TdR concentrations. Two types of DNA histogram analyses were performed on unperturbed and /sup 3/H)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 G0 + G1, S and G2 + M. The results showed that (a) /sup 3/H-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, (b) the increase in the proportion of 4C cells represented a block in G2 and (c) the relative increase in the percentage of 4C cells was proportional to /sup 3/H-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 /sup 3/H-TdR is used to assay substances affecting lymphocyte proliferation or in the estimation of cell cycle time.

• OSTI ID: 5440615
• Journal Information: Cytometry (Baltimore); (United States), Journal Name: Cytometry (Baltimore); (United States) Vol. 1:1; ISSN CYTOD
• Country of Publication: United States
• Language: English

Related Subjects

550301 -- Cytology-- Tracer Techniques
560122* -- Radiation Effects on Cells-- Internal Source-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AGGLUTININS
ANIMAL CELLS
ANTIBODIES
AZINES
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BIOLOGICAL RADIATION EFFECTS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
CELL CYCLE
CELL FLOW SYSTEMS
CELL PROLIFERATION
CONNECTIVE TISSUE CELLS
DNA
HEMAGGLUTININS
HETEROCYCLIC COMPOUNDS
LABELLED COMPOUNDS
LABELLING
LEUKOCYTES
LYMPHOCYTES
MATERIALS
MITOGENS
MUCOPROTEINS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHYTOHEMAGGLUTININ
POLYSACCHARIDES
PROTEINS
PYRIMIDINES
RADIATION EFFECTS
RIBOSIDES
SACCHARIDES
SOMATIC CELLS
THYMIDINE
TRITIUM COMPOUNDS