The Science

Being part animal hospital and part cryonics facility enables us to perform cryopreservation directly after euthanasia. This allows us to achieve lab-quality cryopreservation for pets.

Cryogenic dewar

WHOLE-BODY CRYOPRESERVATION PROCEDURE

Our procedures can vary greatly depending on species, breed, size, and medical state of pets.

STEP ONE: Stabilization

Our procedure begins only after clinical death of the pet once they've reached the end of their natural lives. In ideal cases, this is directly following euthanasia, avoiding ischemic damage and allowing for lab-quality cryopreservation. To begin the process, we administer multiple medications to aid in the cryopreservation procedure and prevent clotting. We place the pet in an ice-water mixture to start to cool down the body.

STEP TWO: Perfusion

Next we transfer the pet to the surgical area and begin perfusing the brain and body with cryoprotectants, which are a sort of medical-grade anti-freeze. This is done via aortic cannulation using a perfusion circuit. As we continue to cool the pet down, higher concentrations of cryoprotectants are perfused until the concentration is high enough for vitrification, ensuring we form a glass-like substance instead of ice crystals. This is vital to avoid the damage that ice formation causes to cells.

Transport to Cryopets facility at dry ice temperatures (−78.5 °C)*

*ONLY IF PREVIOUS STEPS WERE PERFORMED AT EXTERNAL HOSPITAL

STEP THREE: Slow Cooldown

While it's important to quickly cool the body down initially, at lower temperatures we use slow cooling to prevent thermal stress, mitigating fractures. Depending on the size of the pet, this can take a number of days. This is accomplished in our slow-cooldown box, a large device that injects liquid nitrogen into an enclosed chamber over time, gradually dropping the temperature down to −196 °C.

STEP FOUR: Cryostasis

After slow cooling is completed, pets are transferred out of the slow-cooldown box and safely secured in cryogenic dewars, immersing them in liquid nitrogen. These dewars are periodically topped-off with liquid nitrogen to maintain their temperature, and pets are kept there for the long-term until revival. This means pets will maintain their temperature even if there is a power outage.

Electron Micrographs of Vitrified Brain

These are electron micrographs at random locations within the cerebral cortex of a rabbit brain after being perfused with the M22 cryoprotectant, vitrified, and rewarmed. They demonstrate structural preservation with no sign of ice damage. Cryopets uses the M22 cryoprotectant which has been used to successfully cryopreserve and recover entire animal organs.

Research

We’re pushing the science forward in whole-body cryopreservation. Below are some of our interest areas we’ll be exploring on the research end in the coming years.

CRYOPROTECTANT TOXICITY

Cryoprotectants are used to protect cells, tissues, and organs from the damaging effects of freezing and dehydration, but are, themselves, toxic. Our research will look into engineering new types of cryoprotectants to prevent more damage and allow for easier revival in the future.

INTERMEDIATE TEMPERATURE STORAGE

Storing vitrified pets at -196° C enables safe, affordable long-term storage using liquid nitrogen, not electricity. Intermediate temperature storage, around -130° C, may allow for easier revival due to less fractures, but comes with obstacles in affordability and power failure risk. Our research will look into ways around these obstacles to allow for this option.

EMERGENCY KITS

Although ideal cases begin directly after euthanasia, sometimes sudden and unexpected death occurs. Our research will include sourcing and manufacturing the necessary items to develop emergency stabilization kits, allowing pet owners to prevent as much damage as possible.

REWARMING AND REVIVAL

After ensuring we can get the best possible preservation of our pet's brains and bodies, our next priority is working towards repair and revival. Our research in this area will look into rewarming and re-perfusion protocols for whole bodies and investigating the revival technologies needed for pets cryopreserved today such as conventional cell repair and molecular reconstruction.

STANDARDIZED PROCEDURES

As of today, there are not standardized whole-body cryopreservation procedures that are used across multiple cryonics providers. We aim to develop standardized procedures verified by 3rd party neuroscience foundations and performed only by Doctors of Veterinary Medicine. We aim to carefully log every suspension and have transparent quality assurance.

TISSUE PENETRATION

There are certain types of tissues, such as skeletal muscle tissue and fatty tissues, that don't cryopreserve as well as others. Our research will include isolated experiments in cryopreservation of these tissues as well as applied research in improving procedures for whole-body cryopreservation cases.

SLOW-COOLDOWN BOX

The slow-cooldown box is one of the most important pieces of equipment in the cryopreservation process, but has historically been hacked together in various ways at different cryonics organizations. Our research will look into engineering a medical-grade slow-cooldown box to mitigate fractures in the cooldown process.

SUDDEN DEATH AUGMENT

Although sudden death cases are much more rare in pets than in humans, it would be ideal to be prepared for all possibilities. Our research will look into building what we call a Sudden Death Augment (SDA) which will be a surgically implanted device that could take measures to automatically stabilize and cool down bodies in the event of unexpected death.



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