Research Projects

BRLS’s grant recipients include 21st Century Medicine (Fontana, CA); Critical Care Research (Rancho Cucamonga, CA); Suspended Animation (Rancho Santa Margarita, CA); and Advanced Neural Biosciences (Portland, OR).

21st Century Medicine

Organ TransplantsClosing the Gap

Graph showing shortage of organ transplant donations, relative to need
The Organ Shortage Continues
Based on OPTN data from 1991—2017.
See more information at optn.transplant.hrsa.gov.

For more than four decades BRLS has been funding cryobiological research that would allow for increasingly long-term, low-temperature maintenance and storage of transplantable organs. Too often, vitally-needed transplantable organs from accident victims do not reach patients  in time to save their lives because the organs deteriorate so rapidly. As a result, the demand for transplantable organs greatly exceeds the supply. Just as refrigerators and freezers preserve food, low-temperatures preserve transplantable organs. The lower the temperature, the longer the organs can be preserved. However, since freezing – or more specifically, ice-formation – damages biological tissues, cryoprotectants (anti-freeze solutions) must be developed to prevent ice from forming. 

21st Century Medicine (21CM) is a biotechnology company specializing in the cryopreservation of complex systems.  The company, which is located in a cutting-edge research facility in Fontana, California, is one of the premier cryobiological research laboratories in the world. 21CM has developed several relatively non-toxic cryoprotectant solutions. The word “relatively” is used, because all cryoprotectant solutions have some toxicity. For that reason, cryobiologists try to use just enough cryoprotectant to prevent ice formation, without causing toxic damage to tissue.

Of the 150,000 organ transplants that are needed each year in the United States, only about 35,000 are actually performed.  Many exciting advances are leading to ways to eliminate the organ shortage, but an avalanche of new organs will create a new problem – how to manage the volume of resulting transplants.  Even without the added stress of more than quadrupling the number of transplants, transplantation today is a frenetic endeavor: Learjets rushing human hearts to their recipients before their short survival times outside the body expire; surgeons performing transplants in the middle of the night; families on continuous standby, waiting to depart for the hospital at a moment’s notice; organs being transplanted with considerable storage damage; and utilization of poorly matched organs because the only alternative is to discard those organs as their storage times expire.  All of these problems, and more, could be eliminated if organs can be cryopreserved, i.e. if they can be preserved at sub-zero temperatures without time limits.

Cryopreservation, Cryoprotectants and Vitrification: Deep Cooling Without Freezing

Frozen kidney vs. vitrified kidney
Left: A rabbit kidney suspended in a transparent solution of vitrification chemicals, but without vitrification solution inside it, is frozen solid at a temperature of -140°C.
Right: Another kidney with vitrification solution inside it is preserved without freezing at the same temperature of -140°C.
Image courtesy Alcor: https://www.alcor.org/cryomyths.html

Living tissue is mostly made of water.  Adding cryoprotectants to water prevents molecules from gathering together to form ice. Cryoprotectants are chemicals that dissolve in water and lower its melting point.  At -100° C, the molecules become locked in place, forming a non-crystalline, glassy-like material.  This process – by which water becomes solid without freezing – is referred to as vitrification.  Because no ice is formed, biological tissue can be vitrified without structural damage.

21CM has been working on vitrification of various organs, particularly kidneys, with the goal of eventually being able to preserve, or bank, all organ types.

Once most of the water in the organ has been replaced with cryoprotective agents to prevent it from freezing, the next step is to cool it to deep subzero temperatures. This can be done at varying rates, and the effects of these procedures on the organ are then assessed in order to develop protocols for cooling and rewarming that minimize injury.

Indeed, the other side of the vitrification process is rewarming. Using the kidney as an example, cooling down to -45°C before rewarming does not present a challenge because ice cannot form in a kidney at this temperature.  However, when it is cooled to -140°C, ice can form, and significant damage can occur when we try to rewarm the kidney.  To address that problem, rapid warming methods are required. Because the existing technologies are prohibitively expensive, 21CM has been working on rapid warming methods with significantly lower application costs.

Vitrification and Suspended Animation

In the context of cryonics, suspended animation is defined as the demonstrably reversible cessation of all vital functions for an indefinite period of time.  Ultimately, 21CM’s goal is to achieve human suspended animation, arguably one of the most ambitious medical projects ever conceived.  Because suspended animation begins with cryopreservation, the first phase of 21CM’s research focuses on optimizing conditions for vitrifying the brain and other organs, minimizing or eliminating ice formation, and developing reliable and safe methods for measuring vitrification.

Since its founding in 1993, the scientists at 21CM, through funding provided by BRLS, have made history by consistently achieving breakthrough research in a variety of challenging areas.  The research team at 21CM have been granted a number of patents in the areas of cryopreservation, vitrification, and ice-blocker technologies.

Critical Care Research

Ischemia and the Brain

Since 1994, BRLS has funded numerous research projects carried out by Critical Care Research (CCR), which operates a research laboratory in Rancho Cucamonga, California.  The scientists at CCR have made significant progress in protecting against ischemic damage, the tissue damage that normally occurs when blood flow is interrupted.  Disruption of blood flow – which is referred to as ischemia – causes severe cellular metabolic disturbances that often result in irreversible, and eventually fatal damage.  Nowhere is ischemic damage more pronounced than in brain cells, where just a few minutes of blood deprivation can lead to cognitive dysfunction, paralysis and death.

The Implications of Cooling for Modern Surgery

One of the greatest challenges associated with surgical procedures is the risk of neurological impairment induced by ischemia.  When a patient does not survive surgery, it is often because of the occurrence of one or more complications the surgical team did not have enough time to correct before the onset of irreversible brain damage.  The scientists working at CCR have developed medical techniques that could significantly reduce the incidence of surgical complications that lead to neurological impairment.  Using these technologies, surgeons would have several hours to address complications, rather than several minutes as is now the case.  These technologies will also enable more victims of heart attack, stroke and trauma to be revived without permanent brain damage.  Cooling of the brain very soon after an episode of blood flow interruption (i.e. induced hypothermia in post-ischemic events) can prevent brain damage and brain death.

Liquid Ventilation

One of the most significant breakthroughs achieved by the scientists at CCR is an automated Liquid Ventilation system, which can lower whole-body temperatures extremely rapidly by safely introducing cooled liquid perfluorocarbons into the lungs.  The team is currently working to enhance the system so it can be deployed by paramedics  when they respond to emergencies such as automobile accidents, cardiac arrests, strokes and drownings.

Resuscitation Research

CCR is also making great strides in resuscitation research.  Using advanced cooling techniques and other technologies, CCR is currently developing methods to restore life and brain function after varying periods of lack of oxygen and blood flow.  CCR’s development of anti-ischemia drugs, automated induced-hypothermia systems, and advanced cardio-pulmonary rescue systems have the potential to save millions of lives.

Suspended Animation, Inc.

Research

Suspended Animation, Inc. Headquarters

Suspended Animation, Inc. (SA) is a biotechnology company with locations in Rancho Santa Margarita, California and Boynton Beach, Florida.  The company’s research, which is funded through grants provided by BRLS, is focused primarily on developing unique cooling methods to reduce damage to body tissue resulting from loss of blood flow.

When a cardiac arrest, stroke, or trauma occurs, the flow of blood to tissues in the body is interrupted or cut off.  When that happens, oxygen and nutrients are also cut off.  Within minutes, the cells in the tissues have depleted their oxygen reserves, triggering a cascade of biochemical reactions which can ultimately kill the cells.  This is called ischemic damage.

The purpose of the research conducted by SA is to extend the window of time that biological can remain viable and recoverable after cardiac arrest, stroke or trauma. Immediately beginning a treatment protocol that includes rapid cooling of the tissue can help slow, stop, and sometimes even reverse the harmful processes and damaging chemical reactions associated with ischemia.  This allows vital organs to be protected while a patient is being transported to a hospital or other location to receive more advanced medical care.  The additional time provided by this treatment protocol can mean the difference between life and death.

Portable Liquid Ventilation

In collaboration with the scientists at the Critical Care Research laboratory in Rancho Cucamonga, California, SA is designing a portable Liquid Ventilation (LV) unit, which can safely and rapidly cool patients through use of a breathable liquid.  LV units can also be used by first responders and others to treat patients outside of the hospital environment. 

Recovery

SA employs its unique mix of skills and expertise to specialize in the stabilization and transport of cryopreservation patients.

Human organs require immediate cooling and specialized care to stabilize them for transport and to extend  the time window of their viability. Suspended Animation fields teams of highly trained medical professionals with specialized equipment to recover whole body and organ donors throughout the continental United States.  Through its locations in Southern California and South Florida, SA contracts and trains Board certified surgeons, perfusionists and other emergency and critical care professionals. These trained recovery professionals form the 24-Hour emergency response  teams that serve SA’s clients.

Advanced Neural Biosciences

Cryobiology and the Brain

Since 2010, BRLS has funded work conducted by Advanced Neural Biosciences (ANB), a biomedical research company in Portland, Oregon.  ANB, one of the few organizations in the world that conducts research on cryopreservation of the central nervous system, is focused primarily on the following objectives:

  • Perfection of cooling technologies in order to preserve complex brain tissue for the study of neurodegenerative diseases and for advancing/promoting pharmacological drug discovery.

  • Studying damage to the brain caused by the interruption of blood flow (ischemic brain damage), with the goal of developing and improving:
    • Technologies that permit ice-free cryopreservation of the brain after stroke and cardiac arrest.
    • Protocols and chemical compounds that restore optimal blood flow to the brain after stroke and cardiac arrest.

  • Examining the effects of different classes of medications on the cryopreservation of the ischemic brain.