Water transport and cell survival in cryobiological procedures

Abstract

Living cells may be cooled to 77 K (liquid nitrogen) either to destroy them selectively or to store them for long periods. Water transport across the cell membranes during freezing and thawing is a primary factor determining whether the cells survive. These water movements are controlled by phase changes both intracellular and extracellular and by other factors such as the nature of any cryoprotective agent present, and the rates of cooling and thawing. The relation between cooling procedure, water transport and cell survival is discussed. In particular, the crucial rôle of dilution shock is emphasized: this is the damage to cells induced during the dilution that occurs both as ice melts during rewarming and when any cryoprotective additives are removed after thawing. Apart from the usefulness of understanding these processes for maximizing preservation or controlling selective destruction, the diverse responses of cells to different combinations of water transport and temperature changes appear likely to provide basic information on the properties of cell membranes.

Toward Optimal Cryopreservation and Storage for Achievement of High Cell Recovery and Maintenance of Cell Viability and T Cell Functionality

“…The water loss has additional consequences, which are also described as solution effects. 18 The phospholipid membrane can be influenced by dehydration of the cell, leading to an increased sensitivity to mechanical effects. 43,44 Furthermore, the tertiary structure can be damaged due to the high intracellular salt concentration, which can lead to loss of functionality, including denaturation of proteins.…”

“…The loss of water has further consequences, which Farrant described as solution effects. 18 Mazur et al established the two-factor hypothesis wherein they described the optimal freezing rate for successful cryopreservation. 19 Based on these findings, the freezing and thawing processes are now mostly standardized and controlled, for example through the use of freezing containers.…”

Modulation of antifreeze activity and the effect upon post-thaw HepG2 cell viability after cryopreservation

“…Given that cellular damage due to ice recrystallization is directly responsible for the majority of cell death observed during cryopreservation [1,25,28,29,[56][57][58][59]62,74], we explored the ability of the AF(G)Ps in this study to act as cryoprotectants and ultimately ascertain the degree to which cryoinjury can be mitigated by inhibiting ice recrystallization. Previous work by our laboratory has demonstrated that C-AFGPs with ''custom-tailored'' antifreeze activity can function as effective cryoprotectants for a human embryonic liver cell line [15,51,53].…”

Freeze Drying of Red Blood Cells: The Use of Directional Freezing and a New Radio Frequency Lyophilization Device

“…36 RBCs are small cells (6-8 mm diameter) with a defined structure (biconcave) that is linked to their in vivo function as oxygen suppliers. 37 It might be that their water content is low and that in a fast cooling rate (120°C/min-150°C/min) sufficient for water exo-osmosis, combined with the high cell concentration, results in close to 100% survival.…”