TechConnect Innovation Program

KrioBlast™

CELLTRONIX, California, United States

TECHNOLOGY SUMMARY

A novel, robust and scalable platform for hyper-fast cooling that can be used for cryopreservation and cryobanking in medicine, husbandry and wildlife conservation, as well as in a variety of industrial, defense and space applications where cryogenic cooling is a pivotal component.

Primary Application Area: Biotech & Biological Sciences

Technology Development Status: Prototype

Technology Readiness Level: TRL 4

FIGURES OF MERIT

Value Proposition: For each cell type existing technology requires its own freezing method, and often a unique device and a specific costly medium. Achieving high cell viability rate is also a challenge. The KrioBlastTM addresses these issues by unifying the whole process into a single more efficient, automated and scalable platform.

SHOWCASE SUMMARY

Organization Type: Mid-stage Startup (A or B)

Showcase Booth #: 758

Website: www.celltronixbio.com

GOVT/EXTERNAL FUNDING SOURCES

Government Funding/Support to Date: Slow freezing is the major cryopreservation approach. It however requires multi-step protocols, expensive programmable (controlled rate, CR) freezers, and must be tuned to the particular types of cells, tissues and organs.
Ice-free vitrification is the alternative to slow freezing. It is gaining momentum in cryopreservation of embryos, oocytes, ovarian tissues, recently sperm, and some other types of cells. Currently, this approach is impeded by the inability to vitrify big sample volumes at once due to the Leidenfrost effect (LFE), which is the formation of a thermo-insulating vapor coat on the surface of the cryogenically cooled (usually, directly immersed into the liquid nitrogen) sample. It also requires potentially toxic permeable cryopreservatives such as DMSO or ethylene glycol.
As the alternative to these “immersion” methods, we have introduced the KrioBlastTM, a system for hyperfast and scalable vitrfication that is based on spray cooling and completely eliminates LFE.
First, we built a crude manually operated pre-prototype KrioBlast™-1 using own CELLTRONIX recourses. We were able to vitrify large volumes of dilute glycerol solutions (using as cooling rate markers as the relationship % glycerol vs. critical cooling rate is well studied). We then applied for an SBIR Phase I grant. It was granted by the NIH Office of Directors (currently, Office of Research Infrastructure Programs (ORIP), Division of Comparative Medicine), Program Officer: Dr. Miguel Contreras, grant 1R43OD012396-01, total amount $109,000, and it was successfully completed in 2013.
We built a next generation semi-automatic prototype KrioBlast™-2 and reported vitrification of very dilute glycerol-water solutions that accordingly to the literature would represent the cooling rate as high as 600,000 K/min. Thus, we have been able to prove the principle: the KrioBlast™ system is capable to produce extremely high cooling rates.
We then tested the system (the cooler, the container, and the medium) on two clinically important types of cells: human spermatozoa and human embryonic stem cells. The work was done in collaboration with Stem Cell Core of the Sanford-Burnham Medical Research Institute (Prof. Evan Y. Snyder) and UCSD Dept. of Reproductive Medicine (Prof. Sanjay Agarwal). We have reported an excellent survival (around 80-90% of the uncryopresrved cells). However, it also has shown that the system requires some modifications to make it more commercially sound and fully automated. All this will be addressed in the next generation of the system. CELLTRONIX was selected for NIH-sponsored market research. Foresight Science & Technology had conducted market niche analysis for KrioBlast™. We have developed compelling Business Plan; well validated w/ customer and expert surveys. We are planning to apply Phase II SBIR in April, 2014, and estimating our chances as good.
Nevertheless, the Project needs immediate finding as the whole SBIR grant reviewing process will take 9 months. It will give us advantage over our competitors.
We have also submitted a Phase I SBIR grant 1R43DK103578-01 that targets vitrification of hESC-derived insulin-producing; the grant is under NIH review.
We have submitted two non-provisional and one provisional patent applications. Several papers have been published or accepted for publication.

Primary Sources of Funding: Personal Savings, Federal Grant

Looking for: Both Funding and Development Partners