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Trypsinizing and Subculturing Mammalian Cells
05:59

Trypsinizing and Subculturing Mammalian Cells

Published on: June 12, 2008

Trypsinizing and subculturing mammalian cells.

Richard Ricardo1, Katy Phelan

  • 1Molecular Pathology Laboratory, Inc., USA.

Journal of Visualized Experiments : Jove
|December 11, 2008
PubMed
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Subculturing, or passaging, is essential for maintaining cell cultures. This process involves detaching cells and reseeding them to prevent cell death and ensure continued growth.

Area of Science:

  • Cell Biology
  • Tissue Engineering

Background:

  • Cell culture requires regular maintenance to prevent cell death.
  • Confluent cells cease proliferation and eventually die if not passaged.

Purpose of the Study:

  • To describe the essential process of cell subculturing.
  • To outline the critical steps in passaging cell cultures.

Main Methods:

  • Detaching cells from the culture vessel surface using trypsinization or mechanical methods.
  • Reseeding the detached cell suspension into new culture vessels.
  • Monitoring secondary cultures for growth and periodic feeding.

Main Results:

  • Successful subculturing allows for the continuous propagation of cell lines.

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  • Failure to passage leads to decreased mitotic index and cell death.
  • The frequency of passaging is dependent on the specific cell line's growth rate.

    • Conclusions:

    • Cell subculturing is a fundamental technique in cell biology.
    • Proper passaging ensures the viability and proliferation of cells in vitro.
    • Understanding cell growth kinetics is crucial for optimizing subculturing protocols.