Cryopreservation by vitrification: basic thermodynamic principals, methods and devices.

There are 5 basics ways of achieving long-term storage, which ALL essentially lead to vitrification of cells, namely 1. Slow freezing, 2. Equilibrium ice-free vitrification (E-VF) with high concentration of vitrificants and relatively moderate speed of cooling and rewarming, 3. Kinetic intracellular ice-free vitrification (K-VF) with very rapid rates of cooling and rewarming and low to none concentration of exogenous vitrificants, 4. Freeze-drying (lyophilization), and 5. Vacuum/air flow drying at temperatures above zero degree of Celsius (xeropreservation). In this presentation, we will focus on the kinetic vs. equilibrium vitrification. We will compare mechanisms, analyze phase diagrams, emphasize the role of the Leidenfrost effect (LFE) and ways of reducing up to full eliminating LFE. And finally, we will introduce our novel hyper-fast scalable cooling devices KrioBlast™ and VitriPlunger™ and briefly discuss the promising results on K-VF of human sperm, embryonic stem cells and insulin-producing cells using those systems.