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Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation

Nature Protocols volume 3, pages 663–673 (2008)Cite this article

Abstract

Synchronization of cells and nuclei is a powerful technique for the exact study of regulatory mechanisms and for understanding cell cycle events. Counterflow centrifugal elutriation is a biophysical cell separation technique in which cell size and sedimentation density differences of living cells are exploited to isolate subpopulations in various stages of cell cycle. Here, a protocol is described for the separation of phase-enriched subpopulations from exponentially growing Chinese hamster ovary cells at high-resolution power of elutriation. The efficiency of elutriation is confirmed by measuring the DNA content fluorimetrically and by flow cytometry. The resolution power of elutriation is demonstrated by the ability to fractionate nuclei of murine pre-B cells. The installation and elutriation by collecting 16–30 synchronized fractions, including particle size analysis, can be achieved in 4–5 h.

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Figure 1: Schematic view of cell synchronization by centrifugal elutriation.
Figure 2: Counterflow centrifugal elutriation in the JE-6 elutriator rotor.
Figure 3: Synchronization of CHO cells at high resolution of centrifugal elutriation.
Figure 4: Size analysis of synchronized nuclei isolated from murine pre-B cells.

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Acknowledgements

The research related to elutriation was supported by the OTKA grant T042762 (G.B.).

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  1. Department of Microbial Biotechnology and Cell Biology, University of Debrecen, Debrecen, 4010, Hungary

    Gaspar Banfalvi

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Correspondence to Gaspar Banfalvi.

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Banfalvi, G. Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation. Nat Protoc 3, 663–673 (2008). https://doi.org/10.1038/nprot.2008.34

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