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Flow cytometric multiparameter analysis of proliferating cell nuclear antigen/cyclin and Ki-67 antigen: a new view of the cell cycle

Academic Article
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Overview

authors

  • Landberg, G.
  • Tan, Eng
  • Roos, G.

publication date

  • March 1990

journal

  • Experimental Cell Research  Journal

abstract

  • Flow cytometric multiparameter analysis of two proliferation-associated nuclear antigens (proliferating cell nuclear antigen (PCNA)/cyclin and Ki-67) was performed on seven human hematopoietic cell lines. PCNA/cyclin, an S phase-related antigen, was detected using an autoantibody and a fluorescein isothiocyanate-labeled anti-human antibody. The Ki-67 antigen, which in cycling cells is expressed with increasing levels during the S phase with a maximum in the M phase, was detected using a monoclonal antibody and a phycoerythrin-conjugated anti-mouse antibody. In some experiments the PCNA/Ki-67 staining was combined with a DNA stain, 7-amino actinomycin D, and simultaneous detection of the three stains was performed by a single laser flow cytometer. Using this technique four distinct cell populations, representing G1, S, G2, and M, respectively, could be demonstrated in cycling cells on the basis of their PCNA/cyclin and Ki-67 levels. The cell cycle phase specificity could be verified using metaphase (vinblastine, colcemide) and G2 phase (mitoxantrone) blocking agents, as well as by stainings with a mitosis-specific antibody (MPM-2). Also, G0 cells could be discriminated from G1 cells in analysis of a mixture of resting peripheral mononuclear blood cells and a proliferating cell line. This technique can be valuable in detailed cell cycle analysis, since all cell cycle phases can be visualized and calculated using a simple double staining procedure.

subject areas

  • Antigens, Neoplasm
  • Antigens, Surface
  • Cell Cycle
  • Cell Line
  • DNA, Neoplasm
  • Flow Cytometry
  • Humans
  • Ki-67 Antigen
  • Kinetics
  • Nuclear Proteins
  • Proliferating Cell Nuclear Antigen
  • Tumor Cells, Cultured
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Identity

International Standard Serial Number (ISSN)

  • 0014-4827

Digital Object Identifier (DOI)

  • 10.1016/0014-4827(90)90124-s

PubMed ID

  • 1967582
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Additional Document Info

start page

  • 111

end page

  • 118

volume

  • 187

issue

  • 1

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