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A simple, efficient technique for freezing Tetrahymena thermophila

D Cassidy-Hanley1, H R Smith, P J Bruns

  • 1Section of Genetics and Development, Cornell University, Ithaca, New York 14853-2703, USA.

The Journal of Eukaryotic Microbiology
|September 1, 1995
PubMed
Summary
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We developed a simple method for long-term freezing of Tetrahymena thermophila using liquid nitrogen. This technique ensures high recovery of viable cells without specialized equipment, making cryopreservation more accessible.

Area of Science:

  • Cell Biology
  • Cryobiology
  • Microbiology

Background:

  • Long-term preservation of microbial cell lines is crucial for research.
  • Cryopreservation in liquid nitrogen is a common method but requires specialized equipment.
  • Tetrahymena thermophila is a widely used model organism in biological research.

Purpose of the Study:

  • To develop a simple and efficient cryopreservation protocol for Tetrahymena thermophila.
  • To optimize freezing and thawing parameters for maximal cell viability.
  • To assess the applicability of the protocol across different Tetrahymena strains.

Main Methods:

  • Cells were frozen in liquid nitrogen using a simplified protocol.
  • Parameters such as cell physiological state, cryoprotectant exposure time, and freezing/thawing rates were varied.

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  • Viable cell recovery was assessed for multiple Tetrahymena strains.

    • Main Results:

    • The developed technique provides excellent recovery of viable Tetrahymena thermophila cells.
    • The protocol does not necessitate the use of a controlled-rate, low-temperature freezer.
    • Successful cryopreservation was achieved across various tested cell lines.

    Conclusions:

    • A straightforward and effective method for long-term Tetrahymena thermophila cryopreservation in liquid nitrogen has been established.
    • This technique simplifies cell banking and enhances accessibility for researchers.
    • The protocol demonstrates broad applicability for different Tetrahymena strains.