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Related Experiment Videos

Cell synchrony techniques. I. A comparison of methods.

D J Grdina, M L Meistrich, R E Meyn

    Cell and Tissue Kinetics
    |May 1, 1984
    PubMed
    Summary

    Mitotic selection and centrifugal elutriation are effective for synchronizing Chinese hamster ovary (CHO) cells. Cell sorting offers unique isolation capabilities but has limitations in sample size and time for cell cycle studies.

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    Area of Science:

    • Cell Biology
    • Biotechnology

    Background:

    • Achieving synchronized cell populations is crucial for studying cell cycle dynamics.
    • Asynchronous cell cultures present challenges for precise cell cycle analysis.

    Purpose of the Study:

    • To compare the efficacy of different cell synchrony techniques in Chinese hamster ovary (CHO) cells.
    • To evaluate the purity, cell cycle progression, and limitations of various synchronization methods.

    Main Methods:

    • Cell synchrony was induced using mitotic selection, mitotic selection with hydroxyurea, centrifugal elutriation, and EPICS V cell sorting.
    • Cell cycle phase distribution (G1, S, G2+M) was analyzed by flow cytometry.
    • Synchronized cells were replated to assess progression through a second cell cycle.

    Main Results:

    • Mitotic selection yielded pure early G1 phase cells, with cell cycle completion in 12 hours.
    • EPICS V cell sorting produced a pure but heterogeneous G1 population, with cycle completion in 14 hours.
    • Centrifugal elutriation rapidly provided synchronized populations throughout the cell cycle with high purity.

    Conclusions:

    • Mitotic selection and centrifugal elutriation are excellent methods for synchronizing CHO cells, alone or with blocking agents like hydroxyurea.
    • Cell sorting is advantageous for isolating specific cell populations but is limited by sample size and time.
    • Both elutriation and mitotic selection enable robust cell cycle studies in CHO cells.

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