PERSPECTIVE: Temperature-dependent density and thermal expansion of cryoprotective cocktails.

Density is a key thermophysical property, affecting the response of materials to temperature changes in different ways, consistent with the phase of state. In fluids, temperature variation across the domain leads to colder areas being heavier than warmer areas, where buoyancy effects drive fluid flow and thereby increase heat transfer. This phenomenon is known as natural heat convection, which in general is a more efficient heat transfer mechanism than heat conduction in the absence of flow. In solids, where the material is locked in place, colder areas tend to contract while warmer areas tend to expand, leading the material to deform. When this deformation is constrained by the geometry of the domain and/or its container, mechanical stresses develop. This phenomenon is known as thermomechanical stress (or thermal stress), which can lead to structural damage such as fractures. The picture becomes even more complex during vitrification (or glass formation), where the material gradually changes from liquid to an amorphous solid over a significant temperature range. There, due to temperature variation across the domain, fluid mechanics and solid mechanics effects may coexist. It follows that characterization of the density as a function of temperature is crucial for the analyses of thermal, fluid, and mechanical effects during cryopreservation, with the goals of protocol planning, optimization, and preserving structural integrity. For this purpose, the current study focuses on the density of the material and its companion property of thermal expansion. Specifically, this paper reviews literature data on thermal expansion of cryoprotective agents (CPAs), discusses the mathematical relationship between thermal expansion and density, and presents new calculated density data. This study focuses on the CPA cocktails DP6, VS55, M22, and their key ingredients at various concentrations, including DMSO, propylene glycol, and formamide. Data for DP6 combined with a selection of synthetic ice modulators (SIMs) are further presented.