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Related Experiment Videos

A method for freezing sheep lymphocytes prior to cytotoxicity testing

M J Stear, D Allen, R L Spooner

    Tissue Antigens
    |February 1, 1982
    PubMed
    Summary

    A new two-stage freezing method for sheep lymphocytes using high DMSO (17%) achieved over 95% cell viability. This simple, reliable technique preserves lymphocyte antigen expression, making it ideal for microlymphocytotoxicity testing.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Cryopreservation

    Background:

    • Cryopreservation of lymphocytes is crucial for immunological studies.
    • Existing methods may have limitations in viability or antigen preservation.
    • Sheep lymphocytes require optimized freezing protocols for reliable research.

    Purpose of the Study:

    • To adapt and validate a two-stage freezing technique for sheep lymphocytes.
    • To investigate key parameters affecting lymphocyte viability post-thaw.
    • To assess the suitability of the technique for immunological assays.

    Main Methods:

    • Adaptation of a two-stage freezing protocol for ovine lymphocytes.
    • Investigation of lymphocyte concentration, thawing medium composition, and DMSO concentration (up to 17%).

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  • Assessment of cell viability and lymphocyte antigen expression post-thaw.

    • Main Results:

    • The adapted two-stage freezing technique consistently yielded lymphocyte viabilities of ≥95%.
    • A high DMSO concentration (17%) was a key parameter in the modified protocol.
    • The cryopreservation method did not adversely affect lymphocyte antigen expression.

    Conclusions:

    • The developed two-stage freezing technique is simple, reliable, and effective for sheep lymphocytes.
    • High viability and preserved antigen expression make this method suitable for microlymphocytotoxicity tests.
    • This optimized protocol supports advanced immunological research using sheep lymphocytes.