Faltus Milos, Bilavcik Alois, Zamecnik Jiri
Crop Research Institute, Drnovska 507, 16106 Prague, Czech Republic.
Plants (Basel). 2021 Nov 6;10(11):2392. doi: 10.3390/plants10112392.
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. 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 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.
冷冻保护剂(CPA)是当前许多玻璃化方法的重要组成部分。CPA的玻璃化能力会影响玻璃化转变和水结晶发生的概率。在10℃/分钟的标准冷却/升温速率下,利用差示扫描量热法(DSC)评估了两种复合CPA(PVS2和PVS3,常见的植物玻璃化溶液)以及四种单一CPA(乙二醇、二甲基亚砜、甘油和蔗糖,上述PVS的成分)的热特性和玻璃化能力。在所测试的CPA中显示了溶质浓度对其玻璃化能力的影响。定义了玻璃态和/或结晶发生的四个典型浓度区域。我们建议将0.7 g/g的溶质浓度作为通用的玻璃化浓度,其特征是CPA溶液具有实际的玻璃化转变温度(Tg)且水结晶有限。了解CPA的热特性有助于设计具有所需玻璃化能力的新型复合CPA,同时考虑到所使用的冷冻保存方法和冷冻保存样品的特性。