Abstract
With the development of the technique of autoradiography by Howard and Pelc (16), the mammalian cell cycle has become a topic of considerable interest and study. To facilitate this effort, investigators have directed their attention to the development of techniques for use in obtaining cell populations enriched in selected phases of the cell cycle. The technique of mitotic selection (35) described by Terasima and Tolmach in 1961 is one such approach that has enjoyed widespread use and success. By taking advantage of the property of cells from certain cell lines to detach from the surface of culture vessels during mitosis, cell populations enriched in the mitotic phase could be collected through a gentle shaking process. These cells then progress through the cell cycle in a synchronous manner. The major limitation of this procedure, however, is that it is applicable only to selected cell lines that grow in vitro.
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References
Anderson, E. C., Bell, G. I., Peterson, D. F., and Tobey, R. A. Cell growth and division. IV. Determination of volume growth rate and division probability. Biophys. J., 9: 246–263, 1969.
Aqad, S. R., Fox, M, and Winstanley, D. The use of ficoll gradient centrifugation to produce synchrous mouse lymphoma cells. Biochem. Biophys. Res. Comm., 37: 551–558, 1969.
Arndt-Jovin, D. J. and Jovin, T. M. Analysis and sorting of living cells according to deoxyibonucleic acid content. J. Histochem. Cytochem., 25: 585–589, 1977.
Barlogie, B., Spitzer, C., Hart, J. S., Johnston, D. A., Buchner, T., Schumann, J., and Drewinko, B. DNA histogram analysis of human haemopoietic cells. Blood, 48: 245–258, 1976.
Beck, H. P. A new analytical method for determining duration of phases, rates of DNA synthesis and degree of synchronization from flow cytometric data on synchronized cell populations. Cell Tissue Kinet., 11: 139–148, 1978.
Bootsma, D., Budke, L., and Vos, O. Studies on synchronous division of tissue culture cells initiated by excess thymidine. Exp. Cell Res., 33: 301–309, 1964.
Chandler, J. D. Quantum chemistry program exchange. Program 307, Indiana University, Evansville, Ind.
Dean, P. N. Data analysis in cell kinetic research. In: (J. W. Gray and Z. Darzynkiewicz, eds.), Techniques in Cell Cycle Analysis, New Jersey: Humana 1986.
Flow Cytometry and Sorting, (M. Melamed, P. Mullaney, and M. Mendelsohn, eds.) New York: John Wiley, 1979.
Glick, D., von Redlich, D., Yuhos, E. T., and McEwen, C. R. Separation of mast cells by centrifugal elutriation. Exp. Cell Res., 65: 23–26, 1979.
Grabske, R. J. Separating cell populations by elutriation. Fractions, 1: 1–8, 1978.
Gray, J. W. Cell-cycle analysis of perturbed cell population. Computer simulation of sequential DNA distribution. Cell Tissue Kinet., 9: 499–516, 1976.
Gray, J. W., Dolbeare, F., Pallavicini, M., and Vanderlaan, M. Flow cytokinetics. In: (J. W. Gray and Z. Darzynkiewicz, eds.) Techniques in Cell Cycle Analysis. New Jersey: Humana 1986.
Grdina, D. J., Meistrich, M. L., Meyn, R. E., Johnson, T. S., and White, R. A. Cell synchrony techniques. I. A comparison of methods. Cell Tisue Kinet., 17: 223–236, 1984.
Hahn, G. M. State vector description of the proliferation of mammalian cells in tissue culture. I. Exponential growth. Biophys., 6: 275–290, 1966.
Howard, A. and Pelc, S. R. Synthesis of deoxyribonucleic acid in normal and irradiated cells and its relation to chromosome breakage. Heredity (suppl.), 6: 261–273, 1953.
Johnston, D. A., White, R. A., and Barlogie, B. Automatic processing and interpretation of DNA distributions: comparison of several techniques. Comp. Biomed. Res., 11: 393–404, 1978.
Keng, P. C., Li, C. K. N., and Wheeler, K. T. Characterization of the separation properties of the Beckman elutriator system. Cell Biophys., 3: 41–56, 1981.
MacDonald, H. R. and Miller, R. C. Synchronization of mouse L-cells by a velocity sedimentation technique. Biophys. J., 10: 834–842, 1970.
Macdonald, P. D. M. Statistical inference from the fraction labelled mitoses curve. Biometrika, 57: 489–511, 1970.
Macdonald, P. D. M. Age distributions in the general cell kinetic model. In: (A.-J. Valleron and P. D. M. Macdonald, eds.), Biomathematics and Cell Kinetics, Amsterdam: Elsevier/North-Holland Biomedical Press, 1978.
Marquardt, D. W. and Snee, R. D. Ridge regression in practice. Amer. Statistician, 29: 3–30, 1975.
Meistrich, M. L., Meyn, R. E., and Barlogie, B. Synchronization of mouse L-P59 cells by centrifugal elutriation separation. Exp. Cell Res., 105: 169–177, 1977.
Meistrich, M. L. Experimental factors involved in separation by centrifugal elutriation. In: (T. G. Pretlow and T. P. Pretlow, eds.), Cell Separation: Methods and Selected Applications, vol. 2, Florida: Academic, 1983.
Meyn, R. E., Hewitt, R. R., and Humphrey, R. M. Evaluation of S phase synchronization by analysis of DNA replication in S-bromodeoxyuridine. In: (D. M. Prescott, ed.), Methods in Cell Biology, vol. IX, San Francisco: Academic, 1975.
Meyn, R. E., Meistrich, M. L., and White, R. A. Cycle-dependent anticancer drug cytotoxicity in mammalian cells synchronized by centrifugal elutriation. J. Natl. Cancer Inst., 64: 1215–1219, 1980.
Miller, R. G. and Phillips, R. A. Separation of cells by velocity sedimentation. J. Cell. Physiol., 73: 191–202, 1967.
Morris, N. R., Cramer, J. W., and Reno, D. A simple method for concentration of cells in the DNA synthetic period of the mitotic cycle. Exp. Cell. Res., 48: 216–218, 1967.
Rajewsky, M. F. Synchronization in vivo: Kinetics of a malignant cell system following temporary inhibition of DNA synthesis with hydroyurea. Exp. Cell Res., 60: 269–276, 1970.
Romsdahl, M. M. Synchronization of human cell lines with colcemid. Exp. Cell Res., 50: 463–467, 1968.
Roti Roti, J. L. and Dethlefsen, L. A. Matrix simulation of duodenal crypt cell kinetics. I. The steady state. Cell Tissue Kinet., 8: 321–351, 1975.
Sinclair, R. and Bishop, A. H. L. Synchronous culture of strain-L mouse cells. Nature (Lond.), 205: 1272–1273, 1965.
Sinclair, W. K. Hydroxyurea: Effects on Chinese hamster cells grown in culture. Cancer Res., 27: 297–308, 1967.
Stearns, B., Losee, K. A., and Bernsterin, J. Hydroxyurea. A new type of potential antitumor agent. J. Med. Chem., 6: 201, 1963.
Terasima, T. and Tolmach, L. J. Changes in X-ray sensitivity of HeLa cells during the division cycle. Nature, 190: 1210–1211, 1961.
Thames, H. D. and White, R. A. State-vector models of the cell cycle. I. Parametrization and fits to labeled mitoses data. J. Theor. Biol., 67: 733–756, 1977.
Tobey, T. A. and Ley, K. D. Regulation of initiation of DNA synthesis in Chinese hamster cells. I. Production of a stable, reversible G1-arrested population in suspension culture. J. Cell Biol., 46: 151–157, 1970.
White, R. A., Grdina, D. J., Meistrich, M. L., Meyn, R. E., and Johnson, T. S. Comparison of cell synchrony techniques II. Cell Tissue Kinet., 17: 237–245, 1984.
Yen, A., Fried, J., Kitahara, T., Strife, A., and Clarkson, B. D. The kinetic significance of cell size. I. Variation of cell cycle parameters with size measured at mitosis. Exp. Cell Res., 95: 295–302, 1975.
Zeitz, D. S. FPI Analysis. I. Theoretical outline of a new method to analyze time sequences of DNA histograms. Cell Tissue Kinet., 13: 461–471, 1980.
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Grdina, D.J., Meistrich, M.L., Meyn, R.E., Johnson, T.S., White, R.A. (1987). Cell Synchrony Techniques. In: Gray, J.W., Darzynkiewicz, Z. (eds) Techniques in Cell Cycle Analysis. Biological Methods. Humana Press. https://doi.org/10.1007/978-1-60327-406-7_12
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DOI: https://doi.org/10.1007/978-1-60327-406-7_12
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