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|
Cryobiology
Cryobiology: Low
temperature studies of biological systems
By
Krista Nussbaum
Thesis:
Low temperature biology, or cryobiology, has the possibility,
and may very easily and very drastically affect everyones' lives
in the future. Through rapidly approaching cryobiological
medical procedures and techniques, the presence of intense ethical
issues may play an important role in many everyday decisions that
do not now exist.
Outline:
I. Introduction
A. What is cryobiology?
1. definition
2. synonyms
3. general information
II. Cryobiology
Techniques
A. cryosurgery
B. embryo
and gamete preservation
C. tissue
and blood cryopreservation
D. cryotransport
III. Support
of Cryobiology
A. Medical gain
1. techniques
2. knowledge
B. Time gain
1. increase actual length of life
C. having biological children
when you are ready to have them
D. Dispersal of Information
IV. Opposition
to Cryobiology
A. interfering with the
life cycle
B. medical complications
1. physical damage
C. extremely high priced
D. potential law suits
over paternal rights to frozen embryos
E. people may very easily
get carried away with preserving things
V. Conclusion
A. overview of methods
and techniques
B. overview of ethical
issues
C. question for the readers
Introduction
What is cryobiology? It is "the study
of the effects of freezing and low temperatures on living organisms"
(Read, 1999). While the main focus of cryobiology
is on living organisms (as suggested above), cryobiological techniques
have been expanded to include treatment of nonliving things as
well. This addition of non-living objects has expanded the focus
to not only include plants, animals, and humans, but also to include
machinery and other metal objects.
A common synonym of cryobiology is "cryogenics".
According to the New American
Desk Encyclopedia (1989), cryogenics is defined as "the
branch of physics dealing with the behavior of matter at very
low temperatures, and with the production of those temperatures."
As can be seen, cryogenics is a slightly more focused study of
the cryobiological field dealing primarily with production methods
of those freezing temperatures. Another sub-study field, if you
will, is that of cryonics. Cryonics is defined as
the practice of freezing humans who are
not curable by current medical technology, in the hope that ways
may be found to bring them back to life at some future time when
ways of repairing the damage caused by the freezing process have
been developed, as well as cures of the diseases or other causes
of death which necessitated their cryonics suspension (Cryogenics
International, 1999).
As suggested above, cryobiology is primarily
centered on the effects on those living organisms or non-living
objects, although the methods and procedures are also very important.
All around the world new and improved techniques are being tested
to make this a better medical and preservational procedure. Not
only are the different techniques and procedures getting increased
attention, but also many accompanying controversial issues are
as well. Low temperature biology, or cryobiology, has the possibility
to very easily and very drastically affect everyoneís lives
in the future. Through rapidly approaching cryobiological medical
procedures and techniques, the presence of intense ethical issues
may play an important role in many everyday decisions that do
not now exist.
Cryobiology
Techniques
There are many different cryobiological
techniques that exist and are being used today. Such techniques
include cryosurgery, embryo and gamete preservation, tissue preservation
and transplantation, blood and blood product preservation, and
cryotransport. Some of these have been around for so long that
they are now considered commonplace medical procedures, while
others are still considered experimental in the medical world.
Cryosurgery
Cryosurgery, a technique that has been
in operation for years, still has some aspects that are being
extensively tested. Much of the questions and continual testing
revolves around the injury or infection that needs to be treated
and hopefully cured through cryosurgery. Many of these uses of
cryosurgery are widely accepted from a medical standpoint while
others are not widely accepted to the point that in some clinics
and medical facilities they are not provided. According to Gage (1992), "commonly accepted uses of
cryosurgery are for the treatment of many types of skin lesions
(including cancer), for benign and dysplastic mucosal lesions,
for uterine cervicitis and intraepithelial neioplasia, and for
cardiac surgery in the treatment of tacchyarrhythmias". Studies
in all of these areas can easily be found and researched.
Cryosurgery involves different techniques
in which selective necrosis of tissues is achieved, or hoped to
be achieved, through freezing at extremely low temperatures. The
achievement of these low temperatures is through a very precise
cooling sequence, using a specific solution called cryogen. Each
of the separate techniques involves its own specific procedure
in order for the surgery to be successful. Also the goal of treatment
is a determining factor on which cryosurgical technique is used.
According to Kuflik (1994), the three major
goals of treatment to choose from are improvement, cure, or palliation.
Once these initial factors are chosen, the technique to appropriately
achieve the goals is used.
One of these cryosurgical techniques uses
a dipstick apparatus. The dipstick, is simply a cotton-tipped
applicator that is dipped into liquid nitrogen and applied to
a lesion until adequate necrosis has occurred. Another technique
involves an open-spray apparatus by which a fine spray of cryogen
is applied to the lesion. Solidified carbon dioxide can also be
applied directly to the skin with a mix of acetone to treat certain
types of acne. This type of treatment is called slush therapy.
Perhaps the most popular technique used in cryosurgery is the
cryoprobe. This technique involves a precooled metal accessory
that is directly applied to the lesion. The cryoprobe is also
frequently accompanied by liquid nitrogen (Kuflik,
1994).
In most medical facilities, these procedures
are routinely done and have shown both positive and negative results.
Both the positive and the negative results have been reported
on numerous accounts in scientific research papers and journals.
An example of negative research results comes from a study done
by B. Rubinsky, et. al., of the University of California-Berkeley
(1990). This study involves cryosurgical procedures done on the
liver as well as analysis of damage done. They found that the
formation of ice in the hepatic sinusoids expands the sinusoids,
dehydrates the hepatocytes, and then leads to possible structural
damage. This particular study shows that not all cryosurgical
procedures are effective, at least, without causing irreversible
damage to the operation subject in the process.
An example of positive research results
comes from a study of other surgical techniques in comparison
to cryosurgery, by A. Hohki (Gage, 1992).
In this study the other treatment techniques were laser, ultrasonic
aspiration, microwave surgery, scalpel, and electric scalpel.
The major finding was that while all techniques have advantages
and disadvantages, cryosurgery was the best method for cosmetic
effect and economy. In agreement with these findings,
Emmanuel G. Kuflik, MD (1994) has concluded that cryosurgery
has many added benefits besides just cosmetic and economic advantages.
Examples include, suitability for office, nursing home, or outpatient
facility; the lack of need for general anesthesia; optional need
for local anesthesia; the lack of need for operative suits; the
simplicity and safety of the procedure; the lack of physical restrictions
afterward; availability for pregnant patients; and suitability
for those patients who are fearful of surgical procedures.
Not only has there been research-based
proof that cryosurgery is only positive or only negative, but
also there can easily be a mixture of both. A study by Vellet
AD, et al., (1997) shows that cryosurgery for prostate cancer
can have both positive and negative results. "Hypothermic
injury of the rectal wall and bladder base was demonstrated"
after prostate cancer treatment with cryosurgery. However in addition
to this, "no residual prostatic tissue or tumor was found
on follow-up biopsy".
Embryo
and gamete cryopreservation
Although cryosurgery may easily be the
most frequent cryobiological technique, embryo and gamete preservation
comes in a close second. This particular technique has been used
for many species of animals, starting with the mouse in 1972,
to the goat in 1976, and the human in 1983 (Leibo,
1999). Not only has this technique been used for these types
of animal species, in each it has been a successful operation;
successful meaning that live young have been produced from frozen
embryos. In fact, this type of procedure has been so successful
throughout many species of the animal kingdom, the only species
that has still been unable to undergo this procedure with success
is swine (Leibo, 1999). So many animals from
all different species have been born through this process of embryo
cryopreservation that it can be estimated that tens of thousands
individual organisms have survived the storage in liquid nitrogen
at ñ196 degrees Celsius (Leibo, 1999).
Along the same lines as for cryosurgery,
embryo and gamete cryopreservation involves a very precise and
straightforward method of freezing, unique for each organism.
Also along the same lines, many of the methods involve general
steps in the freezing process. These general steps were outlined
by Leibo in his study of preservation of
mammalian embryos (1999). The first step is to expose the embryo
to some sort of cryoprotective solution, for it has been shown
that this is an essential step in successful cryopreservation.
Also, this step is where a lot of further testing has occurred,
and disagreement has been found, on which cryoprotective solution
is the best. The next step is the first step in the actual cooling
process. In this step, the embryo is cooled to slightly lower
than zero degrees Celsius, followed by a seeding procedure. Seeding
is used for inducing ice formation under controlled conditions.
There is much disagreement on whether seeding is actually necessary,
although most procedures require it. The next step, is to control
the cooling of the embryo to an intermediate subzero temperature.
Many studies have shown successful cryopreservation at temperatures
as low as ñ80 to ñ100 degrees Celsius and temperatures
as high as ñ20 to ñ40 degrees Celsius. As mentioned
before, what is being preserved and who was performing the procedure
would determine this temperature. The final step in the freezing
process involves a very rapid cooling to ñ196 degrees Celsius
for long-term storage. It has been estimated that if the proper
freezing steps have been taken, the embryo can remain "alive"
for thousands of years (Leibo, 1999).
Now that you have successfully cryopreserved
an embryo, you must be able to thaw it out in a proper way too.
In general, the warming and thawing of the frozen embryo is directly
related to the amount of freezing that had initially been done;
this is specific for each organism. After warming and thawing
of the embryo is complete, the cryoprotective solution must be
removed, because in most cases, if handled improperly, this solution
is toxic to the embryo (Leibo, 1999).
A new method of embryo cryopreservation
has become a recent cause for study. Katrina Forest and her colleagues
of the University of Wisconsin have been studying the effects
of the cryoprotective solution on the successful preservation
of embryos, and have come up with some striking possibilities
and observations (Bubble Wrap, 1999).
They have observed that immersion in large quantities of cryoprotective
solution has caused the unsuccessful cryopreservation of many
of the test organisms, for example the hamster. As a result, in
their study, they used a technique in which they dipped a thin
nylon loop into the cryoprotective solution and suspended the
embryo on this thin layer of liquid. This allowed them to simply
dip the loop containing the embryo into liquid nitrogen for instantaneous
freezing. Using this method, they have been the only group to
successfully get a hamster birth from a frozen embryo. They hope
that this method would help improve the successfulness of human
embryo preservations in the future.
As mentioned above, most scientists disagree
upon the ideal makeup of the cryoprotective solution. Traditionally,
the cryoprotective solution that has been used was supposed to
simulate the body fluids of the womb, consisting of a sodium ion-
rich saline solution. Until recently, this was a standard procedure.
A study done by James Stachechki (Boyce ,1999)
points out that maybe the success of any cryopreservation is directly
related to the amount of sodium ions in the cryoprotective solution.
In his study, he tested a new solution containing choline ions
(a less abrasive ingredient) on mouse eggs, and got a survival
rate of 90%. This compares to the 50% survival rate of the use
of conventional saline solution. Numerous other studies on improvements
of the cryoprotective solution have been made and many alternate
choices are now available.
Tissue
and Blood Cryopreservation
Cryobiology technology has recently been
focused and manipulated to aid in the care and well being of ill
patients, in a different way than cryosurgery. Tissues and blood
of the body can be removed, frozen, stored, and then later reintroduced
into the body when they are needed or desired. One particular
use of this technology is done for bone marrow transplants. Until
this technology, these transplants relied on time to be effective,
but now cells can be frozen and stored for years and still be
effective. The Cryobiology Laboratory located at the Fred Hutchinson
Cancer Research Center deals directly with this type of cell storage
(Center News, 1997). According to the
laboratory with this technology, "donor and recipient need
no longer be the same ABO blood type [because] the red blood cells
can be removed from the marrow". In this laboratory, the
cells, stem cells or bone marrow cells, are stored for short-term
in a controlled freezer that can cool at a rate of one degree
per minute, or for long-term in a large, liquid nitrogen freezer
at ñ180 to ñ195 degrees Celsius (Center
News, 1997). In this same article, the laboratory has reported
storage of about 1200 patientsí cells, giving a total of
4500 cell packets that are labeled and ready to use when needed.
Since this article was published, the numbers have most likely
increased.
Another form of tissue preservation is
that of blood. In the same manner, blood or blood products can
be stored in freezers and then used when needed by a patient.
Current technology has designed methods of recycling placentas
and umbilical cords, that would normally be discarded after birth,
to be used as blood and hematopoietic stem cell sources. According
to Lifebank Cryogenics Corporation (1999),
this procedure of saving the placenta and umbilical cord at birth
is a very simple procedure that only takes minutes, and in turn
could possibly save your life or the life of your child. These
cells are used for many possible reasons including stem cell matching
for your child, treatment of life-threatening diseases, or even
gene therapy (Lifebank, 1999). Umbilical
cord blood transplantation also has been shown to be an acceptable
alternative to bone marrow transplantation.
CryoTransport
Cryotransport is the newest procedure done
in cryobiology. As defined earlier cryotransport, also known as
cryonics, is the freezing of the entire body. Cryotransport also
can be the freezing of any organ, for example the brain. This
procedure is also known as neuropreservation. A clear-cut method
of how cryonics is done is not in existence, because every cryonics
company or industry has their own specific procedure. One of these
cryonics companies is the Alcor Life Extension
Foundation. They have not only precise methods of preservation,
but also precise methods of training and requirements for training.
Alcor's main objective is to "give people a plausible, 'if-all-else-fails'
pathway to what they have always wanted, something likely to be
achieved in any case, over the next century: an indefinitely long,
healthy life" (Alcor, 1999).
Alcor's cryotransport procedure follows
these steps when a person who has joined Alcor's Life Plan, dies.
First, a standby team immediately starts cooling the body and
giving intravenous medications. The medications are circulated
through the body by external chest compressions or by a specialized
circulation apparatus. This standby team includes four technicians,
one of whom is certified as an EMT, and another who is an Alcor
physician. From the site of death, the cryotransport patient is
then moved to the site of surgery. Here a solution cooled to around
5 degrees Celsius is circulated through the body. This procedure
is known as "washout", and is performed by a team augmented
by a surgeon. This washout procedure must begin within 30 minutes
after cardiac arrest and end within 2 hours to ensure decreased
probability of brain damage to the patient. Once the body has
been stabilized at 5 degrees Celsius, the body is transported
to Alcor's main medical facility where it undergoes the remaining
steps. Here the body's water is replaced with a cryoprotective
agent (glycerol). Over a period of a few hours the concentration
of the cryoprotective agent is increased from 4% to 75%. Upon
completion of this cryoprotection procedure, the body is then
placed into a dewar. A dewar is essentially a large "thermos
flask", which is filled with liquid nitrogen. Liquid nitrogen
is used because it is relatively inexpensive, but it also requires
no refrigeration, thus cutting down on energy needed. The body
can then be kept, preserved in the dewar, for years (Alcor,
1999).
The resuscitation of preserved people has
not yet been successful, although much testing has been done.
"The main problem is that when cells are frozen, water seeps
out of them and collects between them. As this water turns to
ice, it forms crystals which puncture cell membranes" which
cause irreversible damage (Alcor, 1999).
The Alcor Foundation, as well as their patients, believe that
there is a way in which to revive these preserved people and that
it will be found in the near future. For this reason, Alcor's
preservational procedures are so precise and specific.
Although, there are many other cryonics
companies, Alcor is the largest in the world in both popularity
and number of patients. This is why much of the general cryonics
procedures are based on their research.
Support
of Cryobiology
Cryobiology techniques have achieved much
support, especially in recent years. This support has come from
the public, the industries dealing with cryobiology, and also
from the medical field in general. For the most part, cryobiology
has achieved many things in each of these areas. Not only has
cryobiology given new and effective treatments for patients who
need them, it has also provided a substantial gain of knowledge
in the medical field. And this, in turn, allows industries to
make more money and to grow.
The medical gain of what cryobiology has
put into our grasp is sufficient enough for many people to support
this with not only their money, but with their participation.
New techniques would not be effective or even operational if it
wasn't for the persons using the service, or those having faith
in the possibilities presented by them.
The knowledge gained through these newly
evolved procedures in the medical field has been phenomenal. For
example, the different types of cryoprotective solutions has given
the medical field a better handle on how to properly treat certain
tissues and cells in other areas of medicine. New techniques have
been developed, based on prior cryobiological work, which have
also improved the quality of life, and aided in the treatment
of certain illnesses. Many new transplantation techniques and
processes have been gained. The applications of these processes
are getting well known as cryobiology is being further explored.
Many other medical procedures would not even be possible if it
hadn't been for cryobiology.
For many people, the major "plus"
about cryobiology is its ability to lengthen a person's life.
The Full Length Life Society
(1999) is a prime example of this point of view. They have six
major purposes for supporting cryonic suspension of a person.
The first purpose is to give a child who has "died"
a chance to grow. The Society believes that science is on the
road to major discoveries in child illnesses and the treatment
and cure of them, and for this reason they believe that it is
a gift to the family to put a child in cryonic suspension. The
second purpose is a chance to continue parenthood. This purpose
comes into play when a parent "dies", but is not ready
to go or to be left go. The third purpose is a chance to see great
grandchildren. They point out that the scientific process of increasing
the maximum life span is a very time consuming process, and, at
times, in order to meet great grandchildren, you would have to
undergo cryonic suspension. The fourth purpose is a chance for
an extended life span. Many people, just feel that they are not
ready to leave this world, even though their bodies have given
out. The Society says that by undergoing cryonic suspension, one
can literally lengthen their life span. The fifth purpose is a
chance to be born. By this they actually mean that every person
should have the right to be born into a loving and caring family
and at the right time. Cryobiology has allowed unborn fetuses
be kept under cryopreservation for years, until they are desired
by the parents. The Society also believes that this procedure
can easily take the place of many abortion problems, both physical
and emotional. The sixth, and last, purpose is a chance to be
whole. This purpose is geared towards those who have some sort
of major disability, such as, the loss of a limb, the loss of
eyesight, or being a quadriplegic. Through this purpose, as with
the others, the Society feels that science will eventually come
up with solutions for every medical condition (including the ones
mentioned above), and cryonic suspension will allow these new
technologies to affect everyone's lives.
Another form of support for cryobiology
is the dispersal of cryobiological information. The Society
for Cryobiology is a group with more than 400 members, whose
main goal is to do just that (1999). This organization achieves
this goal in two primary ways. Yearly, a scientific meeting is
held, which focuses specifically on cryobiology and related topics.
Also, the Society for Cryobiology
has its own journal entitled Cryobiology. These types of
informative measures not only inform the public, but also those
who are involved in cryobiological science.
Opposition
to Cryobiology
Not only is there much support of cryobiology,
there is also much opposition. This opposition comes in all shapes
and sizes. Many people oppose cryobiology in some forms, but not
in others. An example of this would be, when a person undergoes
cryosurgery to repair a skin lesion, but then opposes cryonics
of a human being. Not only is this type of point of view common,
it is understandable. The parameters of cryobiology and its different
procedures are so far apart that they are hard to lump together
as being similar.
One of the major oppositions to cryobiology
is one of the same reasons why there is much support for it. This
factor would be the ability to increase the length of a life.
Many persons believe that the length of life that we are destined
to should not be tampered with, and this is exactly the reason
why they oppose cryobiology techniques. The ability to live "forever"
is one of the major points of emphasis by cryonics and other cryo-manipulative
techniques, and this brings about much of the controversy. The
definition of "death" is the backbone of this controversy,
because everyone has their own definition. According to the Alcor Life Extension Foundation (1999), "death"
for the whole body is not what actually happens when a person
is pronounced clinically dead, because the individual cells and
whole organs are still alive. Using this definition of "death",
they have justified cryonics as a life-lengthening process. Those
with other definitions, feel that cryonics does not extend life,
but rather may even extend death. Since a successful method has
not yet been found to resuscitate cryonics patients, persons opposing
this technique believe it is not actually a "life-lengthening"
procedure.
Another very important reason for opposing
cryobiology is the potential damage that may occur. Any type of
cryobiological procedure affects the cells and the surrounding
areas, and, thus, could cause damage. In cryosurgery, damage has
been reported right along with the success of treatment for the
problem, as mentioned earlier. In most cases, this damage can
cause future problems that will need to be treated, producing
a viscous cycle. Damage may also occur in embryo or gamete cryopreservation.
Viable births from cryopreserved embryos are, by far, the minority
of all births (Leibo, 1999). This minority
is because of poor technique, simply inactive embryos, or damage
sustained by cryoprotective solutions or the freezer. Birth defects
have been known to be caused by cryopreservation techniques, and
the long-term implications are still unknown to us. The fact that
no successful resuscitations have been performed on cryonics patients
shows that damage has occurred somewhere along the way, or else
that resuscitation of a frozen human being is, in fact, impossible.
The overall price of most cryobiological
procedures is very expensive for the average person to afford.
At the same time, cryobiological procedures are becoming more
of the "norm" in many medical clinics or facilities.
This has the potential for great financial problems when payment
is due, because most people would not have the money. On the other
hand, this great risk of financial burden on those using cryobiology
has been addressed by some of the cryobiology industries. For
example, Alcor Life Extension Foundation
(1999) has formulated payment plans suitable to those inquiring
their services. Alcor has also provided the "patients"
with a fund known as the Patient Care Fund, which is for the benefit
of the "patient" and their family. It seems that not
only has Alcor made payment plans suitable for the "patient"
and their family, but they have also increased the cost of these
procedures in recent years, by considerable amounts. Alcor, in
specific, claims to be a non-profit organization, while at the
same time is charging a fee for cryonic suspension between $50,000
and $120,000. On top of these prices, a whole-life (insurance)
policy costs around $500 a year, $150 for a sign up fee, and $360
per month for annual dues. These prices, along with normal living
expenses, add up to a lot of money to pay, not to mention that
the annual fees extend through the length of cryonic suspension
(Alcor, 1999). These seemingly large prices,
are, yet, another reason why not everyone is supportive of cryobiology.
Legal issues may also arise from some of
these techniques and procedures. For example, what if a married
couple decides to have embryos cryopreserved for later, and more
convenient, use, but before that time comes, they get a divorce.
This introduces the problem of who gets the embryos that used
to be "theirs", but no longer can be? The custody of
frozen embryos is a growing issue, and really is a legitimate
question. Who is in control of stored, frozen embryos and gametes?
And who gets legal rights to them?
There is also the possibility of people
going to extremes with cryopreservation. By this, I mean that
perhaps people will begin preserving inanimate objects such as
their favorite stuffed animal or their lucky shirt, using up valuable
and expensive time, energy, and space. Perhaps this is a very
far-fetched idea, but it is, in fact, possible. Many other concerns
for the effects of cryobiology and cryopreservation exist, but
all can not be mentioned in one short paper.
Conclusion
As described above, cryobiology can be
many different things, including treatment, preservation, as well
as, controversy. The different cryobiological techniques each
have their own equipment, procedures, and ethical issues associated
with them, yet are so similar. While there is much unrest about
whether cryobiology is justifiable, a personal choice is still
the governing force in the matter. For example, my personal
choice can not be influenced by anyone else's. I believe
that some cryobiological procedures I would have no problem participating
and trusting in (i.e. cryosurgery), and others i would never involve
myself or anyone close to me with (i.e. Cryotransport). The theory
of Cryotransport and embryo cryopreservation have very high expectations
for the technology of the future, and that is too much of a risk
for me to take. I guess you could say that I, personally, just
don't have the faith it takes to accept that these theories will
hold true. I believe that cryobiology does have the possibility
for many important medical contributions, but what are we, as
human beings and the inventors of these technologies, willing
to give up in order to achieve them?
Works Cited
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Other References
Lincoln, Tim. (Aug. 28, 1997). "Cold
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McCaffery, Margo. (March 1999). "Pain
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Menezo, Yves J. R. and Moncef Ben Khalifa.
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21, 1998) "A Frankenstein Experiment." Science.
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Taylor, Michael J., Ph.D. (1977). Cryobiology
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