Vitrification Ability of Combined and Single Cryoprotective Agents

Abstract

Cryoprotective agents (CPA) are an important part of many current vitrification methods. The vitrification ability of CPAs influences the probability of the glass transition and water crystallization occurrence. Thermal characteristics and the vitrification ability of two combined CPAs (PVS2 and PVS3), common plant vitrification solutions, and four single CPAs (ethylene glycol, DMSO, glycerol, and sucrose), the components of the mentioned PVSs, were evaluated utilizing a differential scanning calorimetry (DSC) during standard cooling/warming rates of 10 °C min−1. The effect of solute concentration on their vitrification ability was shown in the CPAs tested. Four typical concentration regions at which the glassy state and/or crystallization occurred were defined. We suggest the solute concentration of 0.7 g g−1 as the universal vitrification concentration, characterized by an actual Tg of CPA solution and limited water crystallization. Knowledge of the thermal properties of CPAs allows the design of new combined CPAs with the required vitrification ability respecting the cryopreservation method used and the characteristics of the cryopreserved sample.

Overcoming Challenges for Shoot Tip Cryopreservation of Root and Tuber Crops

“…Therefore, vitrification is an indispensable status to avoid freezing injuries associated with ice crystal formation in biological materials during cryopreservation. As a consequence of the vitrification process, molecular mobility is also highly restricted, and plant tissues can remain viable during long-term storage at −196 • C [113]. Successful vitrification requires dehydration of cells prior to exposure to LN temperatures by exposing plant tissues to high concentrations of cryoprotectants (both penetrating and nonpenetrating) to increase the viscosity of the cell solution [52,61].…”

Cryopreservation of bioflavonoid-rich plant sources and bioflavonoid-microcapsules: emerging technologies for preserving bioactivity and enhancing nutraceutical applications

“…The concentrations vary though. There are different versions of vitrification techniques like encapsulation-vitrification, and droplet-vitrification ( 14 , 95 ).…”

A comprehensive review and update on human fertility cryopreservation methods and tools

“…The most applied protocol for both oocytes and embryos includes the combination of 15% DMSO, 15% EG, and 0.5 M sucrose at the minimum volume of ≤1 μL ( 17 ). A recent study showed that in the most used CPAs combination (DMSO and sucrose), while DMSO reduces the solute concentration, sucrose higher proportion has a direct effect on rising the Tg value, therefore increasing the sample safe storage temperature ( 55 ). Thus, it is needed to expand the knowledge of each CPAs thermodynamics, in order to find the optimal CPA combination for appropriate vitrification.…”