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Related Experiment Videos

Calfection: a novel gene transfer method for suspension cells.

Jeanette Lindell1, Philippe Girard, Natalie Müller

  • 1Laboratory of Cellular Biotechnology, Swiss Federal Institute of Technology, Lausanne, CH-1015, Switzerland.

Biochimica Et Biophysica Acta
|January 30, 2004
PubMed
Summary

We developed Calfection, a novel gene delivery method for mammalian cells using a simple calcium chloride solution. This efficient technique offers high protein expression and long-term storage stability for transfection cocktails.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Background:

  • Gene delivery to mammalian cells is crucial for research and therapeutics.
  • Existing methods like calcium phosphate coprecipitation have limitations, including instability and filtration issues.

Purpose of the Study:

  • To introduce and optimize a novel, simplified method for gene delivery and protein expression in suspension-cultivated mammalian cells.
  • To evaluate the efficiency, stability, and scalability of the new method compared to existing techniques.

Main Methods:

  • Developed "Calfection," a method involving dilution of plasmid DNA into a calcium chloride solution for cell culture addition.
  • Optimized transfection parameters including cell density, calcium concentration, plasmid DNA amount, and culture pH.

Related Experiment Videos

  • Tested Calfection on suspension-adapted HEK293, BHK 21, and CHO DG 44 cells.

    • Main Results:

    • Highest protein expression achieved at 5x10(5) cells/ml, 9 mM calcium, 5 microg/ml plasmid DNA, and pH 7.6.
    • Calfection demonstrated high efficiency with various suspension cell lines.
    • The transfection cocktail remained stable during long-term storage (-80°C to room temperature) and was filterable without loss of efficiency.

    Conclusions:

    • Calfection provides a robust, stable, and efficient method for gene delivery and protein expression in suspension mammalian cells.
    • The simplicity and stability of Calfection make it suitable for large-scale bioreactor applications and high-throughput screening.
    • Calfection offers advantages over traditional calcium phosphate methods due to its stability and filterability.