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Cell Cycle Synchronization pp 31–52Cite as

Synchronization of Mammalian Cells and Nuclei by Centrifugal Elutriation

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1524))

Abstract

Synchronized populations of large numbers of cells can be obtained by centrifugal elutriation on the basis of sedimentation properties of small round particles, with minimal perturbation of cellular functions. The physical characteristics of cell size and sedimentation velocity are operative in the technique of centrifugal elutriation also known as counterstreaming centrifugation. The elutriator is an advanced device for increasing the sedimentation rate to yield enhanced resolution of cell separation. A random population of cells is introduced into the elutriation chamber of an elutriator rotor running in a specially designed centrifuge. By increasing step-by-step the flow rate of the elutriation fluid, successive populations of relatively homogeneous cell size can be removed from the elutriation chamber and used as synchronized subpopulations. For cell synchronization by centrifugal elutriation, early log S phase cell populations are most suitable where most of the cells are in G1 and S phase (>80 %). Apoptotic cells can be found in the early elutriation fractions belonging to the sub-Go window. Protocols for the synchronization of nuclei of murine pre-B cells and high-resolution centrifugal elutriation of CHO cells are given. The verification of purity and cell cycle positions of cells in elutriated fractions includes the measurement of DNA synthesis by [3H]-thymidine incorporation and DNA content by propidium iodide flow cytometry.

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Acknowledgement

This work was supported by the OTKA grant TO42762 (G.B.).

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Authors and Affiliations

  1. Department of Biotechnology and Microbiology, University of Debrecen, 1 Egyetem Square, Debrecen, 4010, Hungary

    Gaspar Banfalvi

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  1. Gaspar Banfalvi
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Correspondence to Gaspar Banfalvi .

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  1. Department of Biotechnology and Microbiology, University of Debrecen, Debrecen, Hungary

    Gaspar Banfalvi

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Banfalvi, G. (2017). Synchronization of Mammalian Cells and Nuclei by Centrifugal Elutriation. In: Banfalvi, G. (eds) Cell Cycle Synchronization. Methods in Molecular Biology, vol 1524. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6603-5_2

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  • DOI: https://doi.org/10.1007/978-1-4939-6603-5_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6602-8

  • Online ISBN: 978-1-4939-6603-5

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