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Second Thermal and Fluids Engineering  Conference

ISSN: 2379-1748
ISBN: 978-1-56700-430-4

KRIOBLAST: A PROMISING CRYOGENIC PLATFORM FOR KINETIC VITRIFICATION BY HYPERFATST COOLING

Igor I. Katkov
CELLTRONIX, San Diego, CA 92126, USA; Laboratory of Amorphous State, National Research University "Bel-SU", Belgorod, Russia

Vladimir F. Bolyukh
CELLTRONIX, San Diego, CA 92126, USA; National Technical University “KhPI”, Kharkov, Ukraine

Abstract

Cryopreservation and cryobanking are insurmountable parts of many sciences and related industries and technologies such as reproductive biology and animal husbandry, husbandry stem cell research and regenerative medicine and, cancer research and precision medicine. Kinetic (very rapid) vitrification (K-VF) is a promising approach for cryopreservation (CP) of biological materials as it is simple, robust, and can achieve VF for practically any type of cells. Several methods of superfast K-VF, particularly for CP of oocytes, embryos, sperm and human embryonic stem cells have been proposed but practically all of them either require very small (in range 0.5-10 μL) size of the sample or/and cannot avoid the Leidenfrost effect (LFE), which substantially impedes the rate of cooling. Here, we are reporting an entirely novel system is based on hyperfast spray cooling of a hermetic cryocontainer for K-VF of one-two order of magnitude larger (thousands of microliters) samples, which we called KrioBlastTM. The system completely eliminates LFE and a need of potentially toxic and osmotically damaging vitrificants such as dimethyl sulfoxide or ethylene glycol used in the current methods of VF. We have been able to stably (repeatedly) vitrify up to 4,000 μL of 15% glycerol solution (used as a cooling rate marker), which theoretically corresponds to the critical cooling rate up to 10,000 °K/s. This platform can be considered as a step toward the "Unified Cryopreservation Protocol".

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