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S-Phase Synchronized CHO Cells Show Elevated Transfection Efficiency and Expression Using CA/PO4

  • Frédéric Grosjean
  • Pascal Batard
  • Martin Jordan
  • Florian M. Wurm
Chapter
Part of the ESACT Proceedings book series (ESACT, volume 1)

Abstract

Various methods exist to transfect mammalian cells in culture. It is generally accepted that individual methods have to be optimized for each of the cell lines or types used. Despite the use of optimized protocols, significant day-to-day variations in transfection efficiency regularly occur. We postulate that the “status” of cell populations prior to transfection is involved in such variability. This study evaluates standardized transfections done at different phases of the cell cycle. CHO cell synchronization was achieved using mimosine. Our results show that applying the Ca/P04-DNA co-precipitation method at different phases of the cell cycle results in variable expression of GFP. Transfection of synchronized cells, during S-phase, resulted in the highest GFP expression level.

Keywords

calcium phosphate cell cycle analysis cell synchronization Chinese Hamster Ovary cells (CHO) flow cytometry Green Fluorescent Protein (GFP) mimosine transient transfection 

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References

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Copyright information

© Springer Science+Business Media Dordrecht 2001

Authors and Affiliations

  • Frédéric Grosjean
    • 1
  • Pascal Batard
    • 1
  • Martin Jordan
    • 1
  • Florian M. Wurm
    • 1
  1. 1.Laboratoire de Biotechnologie Cellulaire, Institut de Génie ChimiqueEcole Polytechnique Fédérale de LausanneLausanneSwitzerland

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