School of Science, RMIT University, Melbourne, Victoria 3001, Australia.
School of Science, RMIT University, Melbourne, Victoria 3001, Australia.
Biochim Biophys Acta Gen Subj. 2021 Jan;1865(1):129749. doi: 10.1016/j.bbagen.2020.129749. Epub 2020 Sep 25.
Cryopreservation is a key method of preservation of biological material for both medical treatments and conservation of endangered species. In order to avoid cellular damage, cryopreservation relies on the addition of a suitable cryoprotective agent (CPA). However, the toxicity of CPAs is a serious concern and often requires rapid removal on thawing which is time consuming and expensive.
The principles of Cryopreservation are reviewed and recent advances in cryopreservation methods and new CPAs are described. The importance of understanding key biophysical properties to assess the cryoprotective potential of new non-toxic compounds is discussed.
Knowing the biophysical properties of a particular cell type is crucial for developing new cryopreservation protocols. Similarly, understanding how potential CPAs interact with cells is key for optimising protocols. For example, cells with a large osmotically inactive volume may require slower addition of CPAs. Similarly, a cell with low permeability may require a longer incubation time with the CPA to allow adequate penetration. Measuring these properties allows efficient optimisation of cryopreservation protocols.
Understanding the interplay between cells and biophysical properties is important not just for developing new, and better optimised, cryopreservation protocols, but also for broader research into topics such as dehydration and desiccation tolerance, chilling and heat stress, as well as membrane structure and function.
冷冻保存是保存生物材料的关键方法,既用于医疗治疗,也用于保护濒危物种。为了避免细胞损伤,冷冻保存依赖于添加合适的冷冻保护剂(CPA)。然而,CPAs 的毒性是一个严重的问题,通常需要在解冻时迅速去除,这既耗时又昂贵。
本文回顾了冷冻保存的原理,并描述了冷冻保存方法和新的 CPA 的最新进展。讨论了了解关键生物物理特性以评估新的无毒化合物的冷冻保护潜力的重要性。
了解特定细胞类型的生物物理特性对于开发新的冷冻保存方案至关重要。同样,了解潜在的 CPAs 与细胞的相互作用对于优化方案也很关键。例如,具有较大渗透压非活性体积的细胞可能需要较慢地添加 CPAs。同样,渗透率低的细胞可能需要更长的时间与 CPA 孵育,以允许足够的渗透。测量这些特性可以有效地优化冷冻保存方案。
了解细胞和生物物理特性之间的相互作用不仅对于开发新的、更好优化的冷冻保存方案很重要,而且对于更广泛的研究也很重要,如脱水和干燥耐受性、冷藏和热应激、以及膜结构和功能。