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Curve -shift analysis of cryopreserved killer T cell function.

R van Lambalgen, J Farrant, B A Bradley

    Journal of Immunological Methods
    |January 1, 1979
    PubMed
    Summary
    This summary is machine-generated.

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    Cryopreservation of cytotoxic effector cells for cell-mediated lympholysis (CML) tests requires careful optimization of cooling rates. Dead cell debris removal is crucial for accurate functional assessment post-thaw.

    Area of Science:

    • Immunology
    • Cell Biology
    • Cryobiology

    Background:

    • Cell-mediated lympholysis (CML) tests are vital for assessing cytotoxic effector cell function.
    • Cryopreservation is essential for storing these cells but can impact their viability and function.
    • Accurate assessment of functional recovery after thawing is critical for reliable CML test results.

    Purpose of the Study:

    • To investigate optimal cryopreservation and thawing conditions for cytotoxic effector cells used in CML assays.
    • To develop a more accurate method for assessing the functional recovery of frozen-thawed effector cells.
    • To identify factors that inhibit effector cell function after cryopreservation and thawing.

    Main Methods:

    • Cryopreservation and thawing of effector cells using dimethyl sulfoxide (DMSO).

    Related Experiment Videos

  • Analysis of functional activity by comparing dose-response curves of fresh versus frozen-thawed cells.
  • Assessment of the impact of dead cell debris and intact dead cells on lytic ability.
  • Use of Ficoll-Hypaque density gradient centrifugation to remove inhibitory debris.
  • Evaluation of vital dye exclusion (eosin) as an indicator of functional cell recovery.

    • Main Results:

    • Optimal cooling rates for cryopreservation varied but were generally between 0.3–1.0°C/min.
    • Dead cell debris, but not intact dead cells, significantly inhibited the lytic activity of thawed effector cells.
    • Ficoll-Hypaque centrifugation effectively removed inhibitory dead cell debris.
    • Vital dye exclusion (eosin) overestimated the functional capacity of thawed effector cells, as it included both functional and non-functional cells.
    • Target cells preserved before phytohemagglutinin (PHA) treatment showed no functional abnormalities post-thaw.

    Conclusions:

    • Optimized cooling rates and removal of dead cell debris are essential for successful cryopreservation of effector cells for CML tests.
    • Vital dye exclusion assays are unreliable for assessing the functional recovery of cryopreserved effector cells.
    • Accurate functional assessment requires methods that evaluate the entire dose-response curve, not just single-point comparisons.