Synchronization of mammalian cells and nuclei by centrifugal elutriation

Research output: Chapter in Book/Report/Conference proceedingChapter

11 Citations (Scopus)

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%). 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.

Original languageEnglish
Title of host publicationCell Cycle Synchronization
Subtitle of host publicationMethods and Protocols
EditorsGaspar Banfalvi
Pages25-45
Number of pages21
DOIs
Publication statusPublished - Aug 4 2011

Publication series

NameMethods in Molecular Biology
Volume761
ISSN (Print)1064-3745

Keywords

  • Counterstreaming centrifugation
  • cell separation
  • elutriator
  • resolution power
  • velocity sedimentation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Banfalvi, G. (2011). Synchronization of mammalian cells and nuclei by centrifugal elutriation. In G. Banfalvi (Ed.), Cell Cycle Synchronization: Methods and Protocols (pp. 25-45). (Methods in Molecular Biology; Vol. 761). https://doi.org/10.1007/978-1-61779-182-6_2