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Cell-dislodging methods under serum-free conditions

H J Cruz1, E M Dias, J L Moreira

  • 1IBET/ITQB-Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.

Applied Microbiology and Biotechnology
|May 1, 1997
PubMed
Summary
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Researchers explored serum-free cell dislodging alternatives for BHK21 cells. Soybean trypsin inhibitor (STI) effectively inactivated trypsin, maintaining cell growth, viability, and productivity, offering a viable serum-free method.

Area of Science:

  • Biotechnology
  • Cell Culture
  • Bioprocessing

Background:

  • Serum is traditionally used for trypsin inactivation in cell culture.
  • Serum-free media are crucial for biopharmaceutical production and reducing costs.
  • Alternative methods for cell dislodging are needed to eliminate serum use.

Purpose of the Study:

  • To evaluate serum-free alternatives for trypsin inactivation in cell dislodging.
  • To identify optimal conditions for cell detachment and subsequent culture.
  • To assess the impact of alternative methods on cell growth, viability, and productivity.

Main Methods:

  • BHK21 cells producing a fusion protein were cultured in protein-free medium.
  • Tested trypsin inactivation with soybean trypsin inhibitor (STI) vs. cell dissociation solution and dispase.

Related Experiment Videos

  • Evaluated the effect of a centrifugation step in conjunction with each method.
  • Determined optimal trypsin to STI ratio for inactivation.

    • Main Results:

    • Trypsin inactivated with STI, followed by centrifugation, yielded the best cell growth, viability, and adherent fraction.
    • Centrifugation consistently improved results across all tested methods.
    • Optimal trypsin inactivation achieved with a 1:1 ratio of trypsin (0.2% w/v) to STI (1 mg/ml).
    • No toxicity observed with STI at tested concentrations.

    Conclusions:

    • Soybean trypsin inhibitor (STI) provides an effective serum-free alternative for trypsin inactivation.
    • The combination of STI-inactivated trypsin and centrifugation is optimal for BHK21 cell dislodging.
    • This method supports long-term cell culture without compromising growth, viability, or productivity.