Dissolved Gases and Ice Fracturing During the Freezing of a Multicellular Organism: Lessons from Tardigrades

Abstract

Three issues are critical for successful cryopreservation of multicellular material: gases dissolved in liquid, thermal conductivity of the tissue, and localization of microstructures. Here we show that heat distribution is controlled by the gas amount dissolved in liquids and that when changing the liquid into solid, the dissolved gases either form bubbles due to the absence of space in the lattice of solids and/or are migrated toward the concentrated salt and sugar solution at the cost of amount of heat required to be removed to complete a solid-state transition. These factors affect the heat distribution in the organs to be cryopreserved. We show that the gas concentration issue controls fracturing of ice when freezing. There are volumetric changes not only when changing the liquid into solid (volume increases) but also reduction of the volume when reaching lower temperatures (volume decreases). We discuss these issues parallel with observations of the cryosurvivability of multicellular organisms, tardigrades, and discuss their analogy for cryopreservation of large organs.

Open‐source hardware‐ and software‐based cryomicroscopy system for investigation of phase transitions in cryobiological research

“…The type of gas bubble formation was described in Ref. (34). The remaining hyperconcentrated solution was in the space between the growing ice crystals.…”

Gas Permeability of Mold during Freezing Process Alters the Pore Distribution of Gelatin Sponge and Its Bone-Forming Ability

“…In contrast, it is known that water typically traps dissolved air [ 32 , 33 ]. The dissolved air is presumed to be dischargeable from ice [ 33 , 34 ] during ice solidification as a result of the absence of space in ice lattices [ 34 ]. This principle is occasionally noted to prepare clear ice in the industry.…”

Fiber Optic Sensor for Real-time Monitoring of Freezing–Thawing Cycle in Cryosurgery

“…This characteristic tissue "bleaching" is partly attributed to a transient blood vessels constriction and the subsequent reduction of the local blood content [3] and partly to the ice crystal and micro-babbles in the tissues. Work by Kletetschka et al [17] showed that depending on the cell structures, which act as nucleation centers, the super-cooled inter-cellular fluids freeze instantaneously forming small crystalline structures. Furthermore, under these conditions of rapid freezing, dissolved gasses inside the cell cannot escape from solution, becoming trapped in the ice forming microscopic bubbles [18,19].…”