Title : Kinetic (hyperfast and free of permeable cryoprotectants) vitrification: A novel approach for precision medicine cryopreservation and biobanking.
Creation of cryobanks (a.k.a. biobanking) is an insurmontabel part of precision medicine. Still, development of an efficient, fast, and robust technology remains a challenge. Cryopreservation (CP) of cells by ultra-fast (tens of thousands C/min) kinetic vitrification (K-VF) of small samples without additional thickeners and ice-blocking vitrification agents (VFAs) had been the first method of cryopreservation back in the 1940s. The mainstream of CP later shifted towards slow freezing (SF). Equilibrium vitrification (E-VF) with very high concentrations of VFAs and relatively moderate rates of cooling and warming (reported in 1980s) turned attention back to VF due it seemingly simple methodology. It however soon became clear that E-VF had had its own substantial limitations due to very high toxicity and osmotic damage produced by VFAs per se. As we pointed out in 2012, the wave has been turning back toward K-VF, especially in the fields of CP of reproductive and germ cells. In this presentation, we touch the basic distinctions between the 3 major ways of CP, with specific emphasis to the means, methods and equipment, as well as pitfalls of the current approaches to K-VF, such as Leidenfrost effect (LFE) and restrictions of the sample size.
A short video demonstrates an entirely new platform for hyper-fast cooling (hundreds of thousands C/min) KrioBlast™ developed by CELLTRONIX, which allows to vitrify relatively large samples (up to 4,000 mcl at the day of reporting, can be further scaled up) practically without a need for potentially damaging vitrificants such as DMSO or propylene glycol, traditionally used nowadays for VF.
The system completely eliminates LFE and a need of potentially toxic and osmotically damaging permeating vitrificants (a.k.a. cryoprotectants) such as DMSO or ethylene glycol used in the current methods of VF. We have been able to vitrify12% and in some case as low as 7% glycerol solutions as a cooling rate markers, which theoretically corresponds to the critical cooling rate up to 1,600,000 C/min or higher. This can be considered as a step toward the “Unified Cryopreservation Protocol”. Preliminary results that demonstrate excellent of survival of human embryonic stem cells and sperm vitrified with KrioBlast™ are also discussed.
Audience Take Away:
• Explain how the audience will be able to use what they learn?
IK: all, who is the dealing with cryopreservation and biobanking will learn about fundamentals of cryobiology and our new and exciting technology of kinetic vitrification.
• Who particularly might be interested in your technology?
IK: both the potential customers and instrumentation companies that can invest to R & D and will have in the future a commercially valuable product.