Alcor Position Statement on Brain Preservation Foundation Prize

From Alcor President, Max More
March 13, 2018

In December 2015, 21st Century Medicine, Inc. published peer-reviewed results of a new cryobiological and neurobiological technique, aldehyde-stabilized cryopreservation (ASC) that provides strong proof that brains can be preserved well enough at cryogenic temperatures for neural connectivity (the connectome) to be completely visualized. This week the Brain Preservation Foundation (BPF), after independent evaluation by neuroscientists and Dr. Ken Hayworth, President of the BPF, awarded The Large Mammal Brain Preservation Prize to 21st Century Medicine based on these results. In 2016 the company had been previously awarded the Small Mammal Brain Preservation Prize for work using the same technology.

Many people are wondering whether Alcor plans to adopt the “Aldehyde-Stabilized Cryopreservation” (ASC) protocol used to win the prize and what the win means for cryonics in practice. Alcor’s position is as follows:

We are pleased that vitrification, the same basic approach that Alcor Life Extension Foundation has utilized since 2001, is finally being recognized by the scientific mainstream as able to eliminate ice damage in the brain during cryopreservation. Alcor first published results showing this in 2004. The technology and solutions that Alcor currently uses for vitrification (a technology from mainstream organ banking research) were actually developed by the same company that developed ASC and has now won both the Small Mammal and Large Mammal Brain Preservation Prize.

ASC under the name “fixation and vitrification” was first proposed for cryonics use in 1986. ASC enables excellent visualization of cellular structure – which was the objective that had to be met to win the prizes – and shows that brains can be preserved well enough at low temperature for neural connectivity to be shown to be preserved. Current brain vitrification methods without fixation lead to dehydration. Dehydration has effects on tissue contrast that make it difficult to see whether the connectome is preserved or not with electron microscopy. That does not mean that dehydration is especially damaging, nor that fixation with toxic aldehyde does less damage. In fact, the M22 vitrification solution used in current brain vitrification technology is believed to be relatively gentle to molecules because it preserves cell viability in other contexts, while still giving structural preservation that is impressive when it is possible to see it. For example, note the synapses visible in the images at the bottom of this page.

While ASC produces clearer images than current methods of vitrification without fixation, it does so at the expense of being toxic to the biological machinery of life by wreaking havoc on a molecular scale. Chemical fixation results in chemical changes (the same as embalming) that are extreme and difficult to evaluate in the absence of at least residual viability. Certainly, fixation is likely to be much harder to reverse so as to restore biological viability as compared to vitrification without fixation. Fixation is also known to increase freezing damage if cryoprotectant penetration is inadequate, further adding to the risk of using fixation under non-ideal conditions that are common in cryonics. Another reason for lack of interest in pursuing this approach is that it is a research dead end on the road to developing reversible tissue preservation in the nearer future.

Alcor looks forward to continued research in ASC and continued improvement in conventional vitrification technology to reduce cryoprotectant toxicity and tissue dehydration. We are especially interested in utilizing blood-brain barrier opening technology such as was used to win the prize.
It may remain unclear to some whether this research and associated prizes show whether ASC or current vitrification without pre-fixation is more likely to preserve cell structures and molecular structures necessary for memory and personal identity. What we can note is that Robert McIntyre, the lead researcher on ASC at 21st Century Medicine, made a point during his presentation at the Alcor 2015 Conference of recommending against adoption of ASC in cryonics at this time.

For cryonics under ideal conditions, the damage that still requires future repair is now more subtle than freezing damage. That damage is believed to be chiefly cryoprotectant toxicity and associated tissue dehydration. It’s time for cryonics debate to move past ill-informed beliefs of “cells bursting.”
This is a groundbreaking result that further strengthens the already strong case that medical biostasis now clearly warrants mainstream scientific discussion, evaluation, and focus.

For a more detailed statement, and one that Alcor endorses, see:
http://www.evidencebasedcryonics.org/2018/03/13/aldehyde-stabilized-large-brain-cryopreservation/
http://www.evidencebasedcryonics.org/media/LBASC.pdf

Corrected analysis of CT scan data show vastly better cryoprotection of Kim Suozzi’s brain

Alcor has been doing CT scans of some patients for the last few years. When we started the project, we did not know how to properly calibrate the output of the scans. This, along with a mistaken assumption, led to a most unfortunately pessimistic assessment of the degree of cryoprotection of Kim Suozzi, the 23-year victim of brain cancer. Now that Alcor has more experience with CT scan data and has created correct calibration standards, we have re-analyzed that report. It turns out that cryoprotection was vastly better than originally reported.

For details see Correction to A-2643 report:

http://www.alcor.org/Library/html/CorrigendumA2643.html

Alcor Position Statement on Brain Preservation Foundation Prize

From Alcor President, Max More
February 12, 2016

In December 2015, 21st Century Medicine, Inc. published peer-reviewed results of a new cryobiological and neurobiological technique, aldehyde-stabilized cryopreservation (ASC) that provides strong proof that brains can be preserved well enough at cryogenic temperatures for neural connectivity (the connectome) to be completely visualized. And this week the Brain Preservation Foundation (BPF), after independent evaluation by neuroscientists Dr. Sebastian Seung, Professor at Princeton, and Dr. Ken Hayworth, President of the BPF, awarded The Small Mammal Brain Preservation Prize to 21st Century Medicine based on these results.

The BPF press release says: “it is the first demonstration that near-perfect, long-term structural preservation of an intact mammalian brain is achievable, thus directly answering what has been a main scientific criticism against cryonics.”

Many people are wondering whether Alcor plans to adopt the “Aldehyde-Stabilized Cryopreservation” (ASC) protocol used to win the prize and what the win means for cryonics in practice. Alcor’s position is as follows:

We are pleased that vitrification, the same basic approach that Alcor Life Extension Foundation has utilized since 2001, is finally being recognized by the scientific mainstream as able to eliminate ice damage in the brain. Alcor first published results showing this in 2004. The technology and solutions that Alcor uses for vitrification (a technology from mainstream organ banking research) were actually developed by the same company that developed ASC and has now won the Brain Preservation Prize.

ASC under the name “fixation and vitrification” was first proposed for cryonics use in 1986. ASC enables excellent visualization of cellular structure – which was the objective that had to be met to win the prize – and shows that brains can be preserved well enough at low temperature for neural connectivity to be shown to be preserved. Current brain vitrification methods without fixation lead to dehydration. Dehydration has effects on tissue contrast that make it difficult to see whether the connectome is preserved or not with electron microscopy. That does not mean that dehydration is especially damaging, nor that fixation with toxic aldehyde does less damage. In fact, the M22 vitrification solution used in current brain vitrification technology is believed to be relatively gentle to molecules because it preserves cell viability in other contexts, while still giving structural preservation that is impressive when it is possible to see it. For example, note the synapses visible in the images at the bottom of this page.

While ASC produces clearer images than current methods of vitrification without fixation, it does so at the expense of being toxic to the biological machinery of life by wreaking havoc on a molecular scale. Chemical fixation results in chemical changes (the same as embalming) that are extreme and difficult to evaluate in the absence of at least residual viability. Certainly, fixation is likely to be much harder to reverse so as to restore biological viability as compared to vitrification without fixation. Fixation is also known to increase freezing damage if cryoprotectant penetration is inadequate, further adding to the risk of using fixation under non-ideal conditions that are common in cryonics. Another reason for lack of interest in pursuing this approach is that it is a research dead end on the road to developing reversible tissue preservation in the nearer future.

Alcor looks forward to continued research in ASC and continued improvement in conventional vitrification technology to reduce cryoprotectant toxicity and tissue dehydration. We are especially interested in utilizing blood-brain barrier opening technology such as was used to win the prize (but which pre-dated work on ASC).

It may remain unclear to many whether this research result shows whether ASC or current vitrification without pre-fixation is more likely to preserve cell structures and molecular structures necessary for memory and personal identity. What we can note is that Robert McIntyre, the lead researcher on ASC at 21st Century Medicine, made a point during his presentation at the Alcor 2015 Conference of recommending against adoption of ASC in cryonics at this time.

For cryonics under ideal conditions, the damage that still requires future repair is now more subtle than freezing damage. That damage is believed to be chiefly cryoprotectant toxicity and associated tissue dehydration. It’s time for cryonics debate to move past ill-informed beliefs of “cells bursting.”

This is a groundbreaking result that further strengthens the already strong case that medical biostasis now clearly warrants mainstream scientific discussion, evaluation, and focus.

For a more detailed statement, and one that Alcor endorses, see:
http://www.evidencebasedcryonics.org/media/MBPP.pdf

Applied Cryobiology – Scientific Symposium on Cryonics

1st to 3rd October 2010 in Goslar, Germany
The German Society for Applied Biostasis (DGAB) organizes its first scientific symposium. It has the title “Applied Cryobiology – Scientific Symposium on Cryonics” and will be held in autumn 2010.

The DGAB was established in May 2006. Aim of the association is the promotion of research and application of biostasis methods, especially of cryonics, for life-extension.

The symposium is a pioneering event. Its aim is to emphasize the scientific fundamentals of cryonics. Scientists from universities and cryonic research-facilities all over the world present the state of the art of science.

During the symposium the “Robert-Ettinger-Medal” for outstanding merits in the field of cryonics will be awarded for the first time. Robert Ettinger is regarded to be the “father of cryonics”. He established the Cryonics Institute in Michigan (USA) and is known to be a pioneer of transhumanism. His books “The Prospect of Immortality” and “Man into Superman” are essential works in the fields of cryonics and transhumanism.

The symposium “Applied Cryobiology” will be held at the UNESCO World Heritage Site Goslar, Germany, from 1st to 3rd October. For more information contact Homepage: http://symposium2010.biostase.de

M22 Implementation

Alcor has implemented a new whole-body cryopreservation procedure which uses a new cryoprotectant solution, M22, licensed from 21st Century Medicine. The new cryoprotectant is perfused throughout the entire body using the same procedures required for glycerol cryoprotection; but while it cryoprotects the torso, arms, and legs, we are confident that it is also vitrifying the brain. The main difference, beyond the composition of the solution, is that neurovitrification can be done without separating the brain from the rest of the body.

This new whole-body procedure will now be standard for all members currently signed up as whole body, as well as for those whole body members who have executed an open option contract.

The important thing to realize about this process is what it cannot do. This is not whole-body vitrification, in the sense that the entire body is vitrified. Significant portions of the body do not cryoprotect sufficiently to vitrify, mostly because things like fatty tissue and skeletal muscle are not well-vascularized. This new procedure may not be as optimum for brain cryoprotection as a simple neuro procedure would be, because of the somewhat longer cryoprotection and cooling times. Conversely, compromises needed to protect the brain may lead to under-cryoprotecting the body, in less ideal cases. But the good thing to remember is that brain vitrification is now available to those who wish to remain strictly whole-body, and that the entire body is cryoprotected intact.

A significant amount of engineering work was done here at Alcor to ensure the proper use of M22. A new circulating chiller and perfusion circuit were needed, as were controlled-temperature enclosures for both the patient and the perfusion circuit to allow safe perfusion at temperatures well below the freezing point of water. The patient enclosure allows further cooling after cryoprotection, to at least -30 or -40 Celsius, reducing the hazards of temperature fluctuations while moving the patient into the next stage of cooling. The credit for this engineering work belongs to Hugh Hixon and Tanya Jones, who have done a wonderful job implementing the new procedure.

Those interested in neurovitrification and whole body cryoprotection will no longer have to face the sorts of compromises they did in the past, and we believe this is a significant advance in our capabilities. For those who would like to know more about M22, the composition and effects have been published in the scientific literature listed below [reference 1]. Electron micrographs of the ultrastructure of brain tissue vitrified with M22 have also been published [reference 2] and are available on the Alcor website at http://alcor.org/library/alcor-new-york-academy-of-sciences-paper/.

Further information and micrographs explaining the switch to M22 technology for all Alcor cases can be found on our website at http://www.alcor.org/library/new-cryopreservation-technology/.

As explained in reference 1, M22 has been shown to be compatible with high viability of tissue slices and with consistent survival of kidneys after transplantation. This allows the unprecedented vitrification of the human brain within the intact human body using a solution that is in principle capable of preserving tissue viability as measured by present-day methods. However, to be sure that the brain will vitrify in compromised human patients, Alcor currently must perfuse M22 for longer periods of time than those shown to preserve viability in model systems. In addition, M22 has not yet been shown to preserve the viability of the brain in model system studies.

Nevertheless, the use of M22 allows Alcor to come closer than ever to achieving the goal of in situ brain vitrification using perfusion conditions that preserve tissue viability by current standards. It keeps Alcor on the road to the possible, eventual attainment of this goal. [SVS/TJ]

(1) Fahy GM, Wowk B, Wu J, Phan J, Rasch C, Chang A, Zendejas E. Cryopreservation of organs by vitrification: perspectives and recent advances. Cryobiology. (2004) Apr;48(2):157-78. http://www.21cm.com/pdfs/cryopreservation_advances.pdf

(2) Lemler J, Harris SB, Platt C, Huffman TM. The arrest of biological time as a bridge to engineered negligible senescence. Ann N Y Acad Sci. (2004) Jun;1019:559-63. Review. http://alcor.org/library/alcor-new-york-academy-of-sciences-paper/

Progress Toward Intermediate Temperature Storage

In the previous Alcor News we reported that Alcor has acquired the prototype of an Intermediate Temperature Storage (ITS) device developed by Brian Wowk of 21st Century Medicine. (For an explanation of why ITS is desirable, please check the explanatory section in Alcor News #13 dated July 1st, 2003.)

Our new lab assistant, Todd Huffman, has been studying Brian Wowk’s design and has visited 21st Century Medicine to discuss reliability issues and possible modifications. Since the ITS pod will be cooled by positioning it above a pool of liquid nitrogen inside a Dewar, we have been debating which type of Dewar to use. An off-the-shelf industrial design is available, large enough to contain seven ITS pods (one in the center and six around it, in a hexagonal pattern). However, our proprietary “bigfoot” Dewar design is much taller, only slightly more expensive, and we have had extensive experience operating them over the past decade. A “bigfoot” seems our best option at this point.

In theory, it would be tall enough to contain a stack of three layers of ITS pods, with seven pods in each layer. The problem is that a tall Dewar will allow a more severe temperature gradient. In other words, the pods at the top will tend to be warmer than the pods at the bottom, and ITS requires that the temperature should be controlled with some precision.

One way around the problem is to use an internal framework of metal such as aluminum, which is a good conductor of heat and could minimize the temperature gradient. Another possibility is to fill the lower part of the Dewar with conventional neuropatients fully immersed in liquid nitrogen, with a single layer of ITS pods above them at the top. The disadvantage of this configuration is that we would have to modify our standard neuropatient containment shell. Also the ITS pods would get in the way during insertion or removal of neuropatients.

Another issue which Todd is investigating is the optimal refill system to maintain the reservoir of liquid nitrogen. A gravity feed would provide the security of constant refill without pumps, but would be grossly inefficient since the pipe connecting it with the Dewar cannot be optimally insulated and will promote nitrogen boiloff.

Using a pump for automatic refill sounds intuitively risky, but low-temperature pump design has been perfected in industrial applications, and a Dewar refill pump probably would run only for a few minutes per week. Two pumps could be installed in parallel for redundancy.

Todd has been tabulating every conceivable failure mode, including liquid-nitrogen level sensor malfunction (Dewar will boil dry), failure of temperature sensor inside ITS pod (patient may become too cold or too warm), wire-break failure, pod heater failure, control system failure, and many others.

Probably we will need four months to explore all these failure modes and develop satisfactory solutions, after which the construction and testing of actual patient storage units may take another two months. This is longer than we would like, but obviously the system must be absolutely reliable before we can offer it as an option to our members.

We can’t predict how much ITS will cost relative to conventional Dewar storage until we have established all the components in the system and have measured the liquid nitrogen boiloff rate.

Intermediate Temperature Storage: A New Era at Alcor

Maintaining patients at an intermediate temperature means keeping them warmer than liquid nitrogen, but cold enough to inhibit biological decay. For years we have wanted to provide this option. On June 14th, we came much closer to our goal.

Why Intermediate Temperature Storage is Necessary

If cryoprotective perfusion is performed successfully with a high terminal concentration, residual amounts of water in solution in the brain tend to solidify instead of forming ice crystals. When we use the vitrification solution which is now standard for all Alcor neuropatients, the entire brain should become a glassy solid as its temperature drops below the “glass transition point” around -125 degrees Celsius.

Traditionally, we have maintained our cryopatients at -196 degrees, the temperature of liquid nitrogen. We use liquid nitrogen because it is cheap, nontoxic, convenient, and requires no refrigeration equipment at our facility. The liquid is “precooled” when it is delivered. Unfortunately, it is colder than we would really like it to be.

When a cryopatient makes the long journey from -125 to -196 degrees, some portions of the brain inevitably tend to cool faster than others. This creates thermal stress which can result in fracturing. We use a “crackphone” to sense and record vibrations which we believe are an accurate indication of fracturing events.

Proponents of nanotechnology believe that fracturing will be relatively easy to repair in the future compared with cellular damage, but still we would like to prevent it. The problem probably can be minimized or even eliminated if the patient isn’t allowed to get so cold, and is held at a temperature just below the glass transition point. In other words, we would like our patients to be cold enough to vitrify, but not so cold that they start to fracture. Unfortunately the only easy way to achieve this has been by using an expensive laboratory freezer–until now.

A New Way to Maintain an Intermediate Temperature

On June 14th, in Rancho Cucamonga, California, biophysicist Brian Wowk of 21st Century Medicine gave a remarkable presentation attended by all Alcor board members and many staff members. Dr. Wowk has developed a simple, reliable design for an intermediate temperature storage device using a heavy-gauge metal container enclosed in a jacket of closed-cell insulating foam fitted with two 2-watt heaters. The insulating jacket is then immersed in liquid nitrogen, and the heaters are run variably by an external controller to maintain the desired temperature inside the metal liner, which conducts heat and minimizes the thermal gradient.

According to Dr. Wowk, seven of his storage devices will fit beside each other within the diameter of a typical “bigfoot” dewar of the design that Alcor uses. The initial cost of building each storage device will be around $2,000, but a greater expense will be incurred in the long term as the heating elements will increase the total boiloff of liquid nitrogen in the enclosing Dewar. Also, because of the foam insulating jacket, each intermediate temperature storage device will occupy a greater volume compared with a standard neuro container. However, we believe that many of our members may feel that a higher payment for longterm care is a very reasonable tradeoff if Dr. Wowk’s design minimizes or eliminates fracturing.

Alcor has purchased Dr. Wowk’s first prototype and will be testing it for reliability and boiloff. After we have the numbers, we will be able to offer intermediate temperature storage probably as an extra-cost option. We can’t estimate the precise cost at this time, but Alcor News will provide additional updates in the future.

This is the most exciting development in cryonics since the advent of vitrification, and we’re especially pleased by its simplicity. We believe there is an excellent chance that this will become the preferred method of patient care at Alcor.