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Related Experiment Videos

Vitrification of human monocytes.

T Takahashi, A Hirsh, E F Erbe

    Cryobiology
    |April 1, 1986
    PubMed
    Summary
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    Cryopreservation of human monocytes using a vitrification solution containing Me2SO, acetamide, propylene glycol, and polyethylene glycol achieved 92% cell recovery. Rapid warming prevented ice formation, preserving monocyte function, while slow warming led to devitrification and cell damage.

    Area of Science:

    • Cryobiology
    • Cellular Biology
    • Immunology

    Background:

    • Cryopreservation is crucial for maintaining cell viability for research and clinical applications.
    • Human monocytes are important immune cells that are challenging to preserve long-term.
    • Vitrification offers a promising alternative to conventional freezing by avoiding ice crystal formation.

    Purpose of the Study:

    • To evaluate the efficacy of a specific vitrification solution for cryopreserving human monocytes.
    • To assess the impact of warming rates on ice formation and cell function after vitrification.
    • To determine the viability and functional recovery of cryopreserved monocytes.

    Main Methods:

    • Human monocytes were isolated using counterflow centrifugal elutriation.

    Related Experiment Videos

  • Cells were suspended in a vitrification solution (20.5% Me2SO, 15.5% acetamide, 10% propylene glycol, 6% polyethylene glycol) and cooled to liquid nitrogen temperature.
  • Vitrified cells were rewarmed at different rates, and viability/functionality was assessed using phagocytosis and chemotaxis assays.
  • Differential scanning calorimetry and freeze-fracture electron microscopy were used to analyze ice formation and devitrification.

    • Main Results:

    • A numerical cell recovery of approximately 92% was achieved.
    • Rapid cooling and warming prevented intracellular and extracellular ice formation, preserving monocyte phagocytic and chemotactic abilities.
    • Devitrification occurred at approximately -70°C during warming, with the extent dependent on warming rate.
    • Slow warming to -70°C resulted in significant intra- and extracellular ice formation, leading to severe loss of cell function.

    Conclusions:

    • The developed vitrification solution effectively cryopreserved human monocytes with high recovery and preserved function when rapid warming protocols were employed.
    • Warming rate is a critical factor in preventing devitrification and maintaining cell viability after vitrification.
    • This method provides a viable strategy for long-term storage and use of functional human monocytes.