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Research article - Peer-reviewed, 2022

Pollen derived macromolecules serve as a new class of ice-nucleating cryoprotectants

Murray, Kathryn A.; Kinney, Nina L. H.; Griffiths, Christopher A.; Hasan, Muhammad; Gibson, Matthew, I; Whale, Thomas F.


Cryopreservation of biological material is vital for existing and emerging biomedical and biotechnological research and related applications, but there remain significant challenges. Cryopreservation of cells in sub-milliliter volumes is difficult because they tend to deeply supercool, favoring lethal intracellular ice formation. Some tree pollens are known to produce polysaccharides capable of nucleating ice at warm sub-zero temperatures. Here we demonstrated that aqueous extractions from European hornbeam pollen (pollen washing water, PWW) increased ice nucleation temperatures in 96-well plates from approximate to - 13 degrees C to approximate to - 7 degrees C. Application of PWW to the cryopreservation of immortalized T-cells in 96-well plates resulted in an increase of post-thaw metabolic activity from 63.9% (95% CI [58.5 to 69.2%]) to 97.4% (95% CI [86.5 to 108.2%]) of unfrozen control. When applied to cryopreservation of immortalized lung carcinoma monolayers, PWW dramatically increased post-thaw metabolic activity, from 1.6% (95% CI [- 6.6 to 9.79%]) to 55.0% (95% CI [41.6 to 68.4%]). In contrast to other ice nucleating agents, PWW is soluble, sterile and has low cytotoxicity meaning it can be readily incorporated into existing cryopreservation procedures. As such, it can be regarded as a unique class of cryoprotectant which acts by inducing ice nucleation at warm temperatures.

Published in

Scientific Reports
2022, volume: 12, number: 1, article number: 12295

Authors' information

Murray, Kathryn A.
University of Warwick
Kinney, Nina L. H.
University of Warwick
Swedish University of Agricultural Sciences, Department of Aquatic Resources
University of Sheffield
Hasan, Muhammad
University of Warwick
Gibson, Matthew
University of Warwick
Whale, Thomas F.
University of Warwick

UKÄ Subject classification

Biochemistry and Molecular Biology

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