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Evaluation of trehalose as a protective agent for isochoric pressure-assisted supercooling of cells.

Author(s) : ZHAO Y., WILLIAM N., PATEL P., WINGNEAN K., ACKER J. P.

Type of article: IJR article

Summary

Effective cryopreservation of biological materials is essential for enabling advances in food storage, biomedical research and clinical applications worldwide. Isochoric pressure-assisted supercooling (Isochoric-PAS) is a promising cryopreservation technique that can preserve biomaterials in a supercooled state without ice damage
by leveraging high hydrostatic pressure. However, exposure to the elevated pressure leads to significant cellular injury. This study investigates trehalose as a protective agent against high-pressure stress during storage of cells with Isochoric-PAS. We hypothesized that although trehalose is generally membrane-impermeant under ambient conditions, high hydrostatic pressure can facilitate its intracellular uptake, enabling stabilization of biomembranes and proteins on both sides of the cell membrane and thereby enhancing cell survival. The experiment was conducted in which Jurkat cells were preserved with trehalose compared with various other cryoprotective agents (CPAs) and subjected to Isochoric-PAS at -2.5 ◦C (31 MPa) and -10 ◦C (111 MPa). Viability and cell size were assessed post-thaw and after a 1-hour recovery. Results show that trehalose at 800 mM yielded superior post-thaw viability (55.0%±1.0% at -10 ◦C) and significantly better recovery after incubation compared with other tested impermeable CPAs. Furthermore, milder freezing conditions (-2.5 ◦C) with lower pressure further improved outcomes. Intracellular trehalose uptake was confirmed and was enhanced under high pressure. This study demonstrates that trehalose is highly effective at mitigating pressure-induced damage, underscoring its potential as a key CPA for optimizing isochoric cryopreservation protocols.

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  • Original title: Evaluation of trehalose as a protective agent for isochoric pressure-assisted supercooling of cells.
  • Record ID : 30035270
  • Languages: English
  • Subject: Technology
  • Source: International Journal of Refrigeration - Revue Internationale du Froid - vol. 189
  • Publication date: 2026/09
  • DOI: http://dx.doi.org/https://doi.org/10.1016/j.ijrefrig.2026.107022

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