Institute of Cell Biophysics of the Russian Academy of Sciences, PSCBR RAS, Pushchino, Moscow Region, 142290, Russia.
The Advanced Research Foundation, Moscow, 119330, Russia.
Cryobiology. 2021 Oct;102:114-120. doi: 10.1016/j.cryobiol.2021.07.004. Epub 2021 Jul 14.
Any biological material contains dissolved gases that affect physical and biological processes associated with cooling and freezing. However, in the cryobiology literature, little attention has been paid to the effect of gasses on cryopreservation. We studied the influence of helium, neon, krypton, xenon, argon, nitrogen, and sulfur hexafluoride on the survivability of HeLa and L929 cell lines during cryopreservation. Saturation of a cell suspension with helium, neon, and sulfur hexafluoride enhanced survival of HeLa and L929 cells after cryopreservation. Helium exerted the most significant effect. For a range of noble gases, the efficiency of the positive effect decreased as the molecular mass of the gas increased. This paper discusses possible mechanisms for the influence of gases on the cryopreservation of biological material. The most probable mechanism is the disruption of the frozen solution structure with gas-filled microbubbles produced during water crystallization. Ultimately, it was concluded that helium and neon can be used to improve methods for cryopreservation of cell suspensions with a low concentration of conventional penetrating cryoprotectants or even without them.
任何生物材料都含有溶解气体,这些气体影响与冷却和冻结相关的物理和生物过程。然而,在低温生物学文献中,很少关注气体对冷冻保存的影响。我们研究了氦、氖、氪、氙、氩、氮和六氟化硫对 HeLa 和 L929 细胞系在冷冻保存过程中的存活率的影响。细胞悬浮液中氦、氖和六氟化硫的饱和提高了冷冻保存后 HeLa 和 L929 细胞的存活率。氦的影响最大。对于一系列稀有气体,随着气体分子量的增加,正效应的效率降低。本文讨论了气体对生物材料冷冻保存影响的可能机制。最可能的机制是在水结晶过程中产生充满气体的微泡,从而破坏冷冻溶液的结构。最终得出的结论是,氦气和氖气可用于改进含有低浓度传统渗透保护剂的细胞悬浮液的冷冻保存方法,甚至可以在没有保护剂的情况下进行。