Natural deep eutectic systems for nature-inspired cryopreservation of cells

Area of Science:

• Biochemistry
• Cryobiology
• Materials Science

Background:

• Natural deep eutectic systems (NADES) show promise as cryoprotective agents (CPAs).
• Optimizing CPA formulations is crucial for effective cell preservation.
• Trehalose-glycerol NADES (T:G) offers a novel approach to cryoprotection.

Purpose of the Study:

• To develop and optimize a trehalose-glycerol NADES-based CPA formulation.
• To investigate the impact of NADES and isoleucine supplementation on cell cryopreservation.
• To elucidate the mechanisms underlying NADES-mediated cryoprotection.

Main Methods:

• Differential scanning calorimetry (DSC) for thermophysical property analysis.
• Raman spectroscopy to study ice formation and hydrogen bonding.
• Cryopreservation of Jurkat cells followed by recovery, apoptosis, and growth assays.

Main Results:

• NADES formulations effectively suppressed ice formation and dehydration.
• Isoleucine supplementation did not alter thermophysical properties but significantly improved cell survival and proliferation.
• Raman spectra and heat maps provided insights into NADES's effect on solution structure.

Conclusions:

• Optimized NADES formulations enhance cell cryopreservation outcomes.
• Isoleucine supplementation is a critical factor for improving post-thaw cell viability.
• Cryoprotective efficacy depends on more than just thermophysical properties, highlighting the role of biological structure stabilization.