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. 1972 Sep 1;54(3):638–645. doi: 10.1083/jcb.54.3.638

A METHOD FOR COMPARING EFFECTS OF DIFFERENT SYNCHRONIZING PROTOCOLS ON MAMMALIAN CELL CYCLE TRAVERSE

The Traverse Perturbation Index

Robert A Tobey 1, Harry A Crissman 1, Paul M Kraemer 1
PMCID: PMC2200279  PMID: 5044762

Abstract

After treatment of Chinese hamster cells (line CHO) with various protocols for synchrony induction, the subsequent ability of cells to traverse the cell cycle (i e., to perform, an essential cell cycle process) has been determined by measurement of the DNA distribution pattern among cells in large populations with the Los Alamos flow microfluorometer In the cultures prepared by the various synchronizing techniques the vast majority of cells traversed the cell cycle in a normal fashion; however, in all cultures examined there remained small subpopulations which, though remaining viable for several days, could not carry out normal traverse. After reversible inhibition of DNA synthesis by means of a double-thymidine blockade, approximately 17% of the cells were unable to complete genome replication. After reversal of G1 arrest resulting from cultivation of cells in isoleucine-deficient medium, 12 4% of the cells commenced synthesis of DNA but were unable to complete the S phase. Cells prepared by mitotic selection yielded a subpopulation (5 5% of the total cells) with a G1 DNA content which remained viable but noncycling for at least 5 days. We propose a term "traverse perturbation index" which is defined as the fraction of cells converted to a noncycle-traversing state as the result of experimental manipulation. A knowledge of the perturbation index will allow direct comparison of effects on cell cycle traverse of various synchrony-induction protocols

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bostock C. J., Prescott D. M., Kirkpatrick J. B. An evaluation of the double thymidine block for synchronizing mammalian cells at the G1-S border. Exp Cell Res. 1971 Sep;68(1):163–168. doi: 10.1016/0014-4827(71)90599-4. [DOI] [PubMed] [Google Scholar]
  2. CULLING C., VASSAR P. Desoxyribose nucleic acid. A fluorescent histochemical technique. Arch Pathol. 1961 Jan;71:76–80. [PubMed] [Google Scholar]
  3. Galavazi G., Bootsma D. Synchronization of mammalian cells in vitro by inhibition of the DNA synthesis. II. Population dynamics. Exp Cell Res. 1966 Feb;41(2):438–451. doi: 10.1016/s0014-4827(66)80150-7. [DOI] [PubMed] [Google Scholar]
  4. Petersen D. F., Tobey R. A., Anderson E. C. Synchronously dividing mammalian cells. Fed Proc. 1969 Nov-Dec;28(6):1771–1779. [PubMed] [Google Scholar]
  5. Studzinski G. P., Lambert W. C. Thymidine as a synchronizing agent. I. Nucleic acid and protein formation in synchronous HeLa cultures treated with excess thymidine. J Cell Physiol. 1969 Apr;73(2):109–117. doi: 10.1002/jcp.1040730204. [DOI] [PubMed] [Google Scholar]
  6. Tobey R. A., Anderson E. C., Petersen D. F. Properties of mitotic cells prepared by mechanically shaking monolayer cultures of Chinese hamster cells. J Cell Physiol. 1967 Aug;70(1):63–68. doi: 10.1002/jcp.1040700109. [DOI] [PubMed] [Google Scholar]
  7. Tobey R. A., Ley K. D. Isoleucine-mediated regulation of genome repliction in various mammalian cell lines. Cancer Res. 1971 Jan;31(1):46–51. [PubMed] [Google Scholar]
  8. Van Dilla M. A., Trujillo T. T., Mullaney P. F., Coulter J. R. Cell microfluorometry: a method for rapid fluorescence measurement. Science. 1969 Mar 14;163(3872):1213–1214. doi: 10.1126/science.163.3872.1213. [DOI] [PubMed] [Google Scholar]

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