Experiments in the field of biotechnology conducted by the USA Apollo and Skylab space probes are described, as well as the joint Soviet-American Apollo-Soyuz Test Project (ASTP). Experiments in electrophoretic separation in space of biological compounds in a liquid medium are detailed. Space processing of vaccines and separation of human and animal cells are described. Methyl-cellulose, a coating for use in electrophoresis was developed. Erythropoietin, which stimulates the formation of red blood corpuscles in bone marrow, was obtained in pure form

Gilyarov, V. N.

English

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NASA TECHNICAL MEMORANDUM NASA TM-76235
CERTAIN PROBLEMS OF SPACE BIOTECHNOLOGY
{NASA-Vi-76235) CERTAIN PROBLEMS OF SPACE
BiUTECHNOLUGY {atiou)al1Lep HCuA0z/nHFA01
Space Adaiuistr atiou) 	CSCL 06C
S80- 31411
Uuclda
3/12 28669
Translation of "Nekotoryye zadachi kosmicheskoy
biotekhnologii",
	 Academy of Sciences,
USSR, Institute of Space Research, Moscow, Report
Pr-356, 1977 , pp 1-13
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NATIONAL AERONAUTICS AND SPACE ADMINISTRATION
WASHINGTON, D.C.	 JUNE 1980
PAGI
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WHO
WCERTAIN PROBLEMS OF SPACE BIOTECHNOLOGY
V. N. Gilyarov
The utilisation of space apparatus for conducting a series of techno-	 jI *
logical processes opens additional new possibilities in the field of biology
and medicine. This is conditioned by: the lack of weightlessness in heat convection
(i.e., the lack of any internal processes of movement in material except for
insignificant convective-like currents due to variation in surface.tension
and non-uniformity of thermal expansion)= the lack of sedimentation and
division by weight not only of solid, liquid, and gaseous products of
reactionf., but also directly in the processes of growth and development of
microorganism and tissue cultures. As a result, the effectiveness and degree
of purity in the division of mixtures during electrophoresis is increased,
which may be used for obtaining pure specimens of albumen, subcellular
particles, and other biological materials and for obtaining vacc4-nes with
a very high degree of purity.
The totality of processes conducted under conditions of space flight
during which qualitative changes take place in a utilized biological material
and as a result of which new medical or biological materials are obtained,
as well as the scientific descriptirn of these processes is the new field
of space technology -- biotechnology.
Scientists abroad had begun developing this new direction as early 	 I4
as the end of the 60's. Included among prospective technological operations
in NASA's plans in the -70's in developing a project for a long-term orbital
station was the process of obtaining vaccines by means of electrophoresis
under conditions of space flight C 'I .
*Numbers in margin indicate pagination in original foreign text.
1
Electrophoresis is the movement of disperse particles found in a
suspended state under the action of an external electrical field. Their
movement is associated with the existence of a double electrical layer on
the boundary with the medium.
I•.ost molecules or organic substances found in aqueous solution of
acid or alkali take on small electrical charges, and move through this
solution under the influence of applied electrical field. since different
molecules move at different speeds, the faster molecules in a mixture of
organic substances, which begin to move from one end of a tube fil. :d with
solution to the other, will go ahead of the slower molecules.
The effect of dividing a homogeneous mixture of biological materials
during flora in a uniform electrical field opens new possibilities for
cytology, immunobiology, and general chemistry. Under earth condi-_ions,
the effectiveness of separation is reduced due to sedimentation and
convective currents.
Llectrophoretic division of biological compounds in a liquid mediwn
was first conducted on the "Apollo-14" spacecraft (1971). These operations
were recorded on film and transmitted by television.
The apparatus for such division consisted of three tubes: 1 - containing
a mixture with red and blue organic dyes; 2 - containing human hemoglobin;
3 - containing DNA from salmon roe, suspended in a dissolved electrolyte
solution (a mixture of water, boric acid, and sodium hydroxide). All these
organic compounds have different molecular weights. 	 j5
The device also includedt a pump with motor, gas separator, fluorescent
lamps, energy source, and thermometer. The weight of the entire device was
2
4.082 kg in a packaging size of 177 x 127 x 101.6 mm.
The power consumed was 30 W from a source of alternating current with
frequency of 400 Hz. While the experiment was being conducted, all equipment
was secured with the aid of straps. The experiment lasted 1 hour. A+l
processes were filmed on color film with the aid of a 70-mm on-board camera.
Each of the three tubes had electrodes built into its ends. The
electrodes were separated from the medium by porous membranes. power was
fed to the electrodes, and the electrolyte circu'ated due to the operation
of the pump which was powered by a motor. A constant current was passed
through the liquid (230 W). Hydrogen and oxygen formed in the solution
were absorbed by gas separators. The fluorescent lamps illuminated the
tubes for photography. Visible light was used for illuminating the dyes
and the hemoglobin, and ultraviolet light has used to illuminate the salmon
roe DI A.
The equipment for electro.horesis on the "rpollo-14" spacecraft was
developed by General Electric under contract with the Center for .anned
Space Flight in roz ston C 2 0 3^
In 1972 on the "Apollo-16" spacecraft, along; with biolo gical experiments
on the study of the influence of individual factors of space flight on
bacteria, fungi, and viruses, cellular elements in the blood of humans and
rabbits were also divided.
The demonstration of electrophoretic separation was intended for
testing technical solutions for the preparation of the process of obtaining
pure specimens of organic substances in space. In particular, this experiment
had in perspective an important practical significance for the preparation
of vaccines.
3
rBy 1974 it became clear that electrophoresis was not only possible 	 f6
under conditions of space flight, but also opened up grew: possibilities
for the purification and separation of biological materials.
Conditions in space create an exceptionally favorable environment for
obtaining vaccines in large quantity and of high quality. In the absence
of gravitational forces, the speed of growth of microorganisms may be
doubled. The processes of fermentation and bacteria culture turn out better
in space than they do on earth. The lack of gravity "unhinders" their
metabolism and facilitates their oxygen supply.
further, virusFs which serve in the prepar-i-tion of vaccines must be
separated from the nutrient medium on which they are grown. weak admixtures
in vaccines prepared on Larth may cause undesirable side effects. The
co.centration of useful agents in these vaccines is also insufficient for
their necessary effectiveness. Also, on Larth, due to convective currents,
it is possible to purify solutions which are subje:.-ed t? an electrical field
either in the form of E. thin film or in porous material, which makes it
impossible to obtain vaccine in large volume L 4 - ? I
On the basis of experiments conducted on the Apollo program and Skylab,
American scientists believe that in space it will be possible to purify the 10
most necessary types of vaccines. •.ereover, they feel th- In one year it
is possible to obtain 1 ton of vaccine (10 types), which would supply the
needs of the entire world, with an expenditure of several million dollars
for obtaining this vaccine. In particular, it is planned that the shuttle
program (multi-use transport space vehicle) and rocket satellites found in
a state close to weihtlessness for a period of 5 - 12 ntin L 81 are to
be used for this purpose.
4
Aside from an economic advantage, the production of vaccines in space
Will make it possible to obtain vaccines of the highest effectiveness in
combatting flu, infections, and catarrhal diseases. Components may be
serarated and concentrated with a high degree of accuracy.
Already now NASA and 1:5A (huropean Space Administration) are working
on developing equipment for static electrophoresis, for measuring the
mobility of material, for flowing materials and isotachophoresis, where
the sample mixture is layered between substances with different ion
properties.
Among the scientific experiments in the joint Soviet-American AST.'
program (1975) was the electrophoretic separation for medical-biological
research: obtaining pure and isolated specimens. The experiment studied
the effectiveness o1 using electrophoretic processes for separating lymphocytes,
enzymes, as well as other components of blood and tissue liquid of man
under conditions of weightlessness. In preparation for the experiment, an
improved method of freezing and subsequent long-term storage (up to 5 years)
of white blood corpuscles (granulocytes) without altering, their immunological
properties was developed C 9, 101 .
The separation of human cells in closed tubes filled with immobile
medium was envisioned. The purpose was to attempt to separate certain types
of cells, which is difficult or completely impossible on it;arth.
In planning the conditions for conducting the experiment, it was
necessary to solve the problems of creating equipment capable of withstanding
acceleration during launch and descent, the problem of removing gas bubbles
from the electrodes and keeping the results of the experiment.
17
5
A device weighing 26 kg was designed by the P•VV company (German
Federal Republic), and the Planck Institute of Biochemistry was in charge
of the experiment
	 9 - 12, .
Four samples were tested in the course of the experiments a mixture of
	
/f3
human and rabbit erythrocytes; cells from rat spleen, cells from rat bone
marrow, and a mixture of lymphoid cells of rats and human erythrocytes.
Three electrophoresis regimes were tested. In the first, the field intensity
comprised 60 w/cm, the speed of pumping; buffer solution -- 0.2?5 ml/sec, and
speed of sample flow -- 5 ml/hr. In the second regime, the speed of sample
flow was reduced to 1 ml/hr= in the third it remained equal to the value
of the second regime, but the field intensity and speed of buffer solution
floe: were reduced by 113.
The quality of separation was improved by ' J 10 timer [ 11 - 15] .
The second experiment consisted of separating from the nutrient mediutu
active kidney cells possessing the property of producing; the enzyme urokinase,
which hinders the formation of blood clots in human blood vessels. irokinase
dissolves the blood cell membranes and is used as an initial raw material for
the creation of effective preparations for treating phlebitis. rokinase is
produced by •v 5,^"p
 
of kidney cells. Their separation in flight with the id of
electrophoresis was implemented as follows: Kidney cells were introduc.d
into one end of a tube filled with water, and then an, electrical field was
created. Due to the different polarization of active and inactive kidney
cells, their speed of i.ovement in the electrical fief in the absence of
gravity is also different, and this leads to their separation 	 1!! .
If this experiment were to be conducted under earth conditions,
gravitation, would cause convective currents, and then all the cells would
6
settle. It was rioted that under conditions of space flight, the cells
excreted 6 - 7 times pore urokinase than on earth. E,oreover, for the
production of considerable quantities of urokinase, a lwr ge amount of
initial material is necessary. '.'hus, obtaining this substance under
conditions of weightlessness is economically expedient.
Separated kidney cells were frozen and returned to earth for .subsequent 	/9
cultivation. These cells are presently being used successfu?l.T for forming
addit'.onal cell structures.
Also, a new coating in the device used for conducting electrophoresis
was developed -- methyl-cellulose. it removes certain electrical changes
in the medium and this gives an even clearer division of the material. The
new coating maintains its properties over a long period of time.
Another candidate for production in space is erythropoietin, which
stimulates the formation of red blood corpuscles in bone narrow. On earth
It cannot be obtained from kidneys in pure form, and the presence of a number
of antitoxins in it causes undesirable side effects [171 .
t. ulti-use transport space vehicles may be used in organizing the production
of eiythropoietin in quantities sufficient not only for practical utilization,
but also for research work. however, obtaining; even small quantities of
this preparation is important and useful.
Thus, certain results of medical-biological experiments of the A6Ti
program may find widespread application in medicine. In particular, a new
method has already been found for storing donor blood without changes in
Immunological properties for transfusions to leukemia patients i, new
composition of storage medium has been developed, which makes operations
for transfusion of tXanulocytes more reliable and accessible. Also as a
7
result of these experiments, the possibility was shown for separating from
the kidney cell culture a cell subpopulation, which effectively produces
urokinase enzyme, which inhibits processes of thrombosis of the cardio-
vascular system.
According to the evaluation of GSA specialists, conducting technological
processes on spAoe vehicles for the purpose of obtaining biological materials
may in the future become one of the main aspects of utilizing space and may 	 j10
create a "market" of 1 billion dollars per year. Listed among the materials which
may possibly be obtained is-an improved serum for use in kidney transplants
and certain hormones C 181 .
A colloquium held at Johnson Center (USA) in Larch of 1 976 was devoted
to questions of producing biological materials in space. The colloquium
was organized by NASA. It ^::.amined the possibilities of conducting medical-
biological research on board the space shuttle and in the "Space lab" block.
A representative of i+ASA, in particular, a,:nounced that on board the space
shuttle and in the Space Lab block, 300 - 400 separate experiments may be
conducted during each flight. Specialists at the colloquium examined the
possibilities of conducting research in the directions of:
1. Studying the behavior of living cells under space conditions,
including the cells of invertebrates. For example, sponge cells produce
substances of an anticareinogenic nature and antibiotic type substances.
It would be important to develop such qualities of cells.
2. Studying the influence of high energy particles, for example, on
tumoral cells. Under conditions of space flight, particles with energy of
up to 10 40  L eV are accessible for experiments (under earth conditions using
accelerators -- no higher than 2 i-,eV)'. 11ice which have been contaminated with
tumoral cells will be irradiated with the indicated energies for a period
of 10 - 30 days.
8
1©
3. Growing anaerobic organisms by obtaining their metabolic products
under conditions of space vacuum and lack of oxygen.
4. Utilizing cosmic ultraviolet radiation for more effective sterilization
of viruses with fewer difficulties than under earth conditions using the 	 jll
same radiation.
5. Genetic engineering. Under conditions of weightlessness by performing
manipulations on genetic systems it will be possible to avoid deviations of
the type of branching genetic chains or bonding of these chains.
6. Obtaining biochemical substances under conditions of weightlessness
with the lack of convection. For example, a hormone with very promising anti-
carcinogenic properties has been discovered. However, under earth conditions
they are obtained only in micrograms with excessively high cost. Under
conditions of weightlessness, perhaps, it will be possible to obtain this
substance in quantities measured in grans.
It was also mentioned at the colloquium that the equipment developed
for obtaining materials of this type under conditions of space flight may
be used later on earth after appropriate modification, which would give a
definite economical advantage C 193 .
questions of utilizing conditions of space flight for industrial
production in space of certain types of unique preparations are still being
discussed to this day. It is not the aerospace organizations of the USA and
the Luropean countries who are enlisted for this purpose. The suggested
problems are being thoroughly exam-A.ned and economically analyzed.
It is assumed that a number of problems in space biotechnoloGy will be
solved in the space shuttle flights during the time of 1979 - 19U2. This
concerns, for example, the question of obtaining most important productst
9
highly pure vaocines, highly effective viral inseuticides, highly effective
!.soenzymes, albumens, hormones, immunoglobulin. The separation of iso-
enzymes may brim a clear profit of 20 - 29%. There are tentative plans
to include on three flights of an orbital laboratory devices for electro-
phoretic separation of preparations of the urokinase and erythropoietin type 1 20-22, .
From an examination of even this far-from-complete list of problems	 /L2
in space biotechnoloiV. the huge signifiearce of their solution is clearly
evident for the purpose of practically obtaining a number of materials
as well as for the further development of scientific research work in this
field.
10
REFERENCES
1. Space 'World, 1971, V,H-1-85, 4-12
2. Space World, 1971, V.H-4-88, 28-30
3. Aviation Week & Space Technology, 1971, V.94, N7,9, 17•-25
4. Spaceflight, 1972, V.14, Nr, 246-253
5. Aviation Week & Space Technology, 1972, V.96,N17, 18-20
6. Aviation Week & Space Technology, 1970, V.92,N25, 23
7. Aviation Week & Space Technology, 1970, V.93,N18, 53
A. Spaceflight, 1974, V.16, N7, 271-272
9. Spaceflight, 1976, V.18,N4, 136
10. Defeuse/Space Business Daily, 1976, V.84, O f 43
11. Raunfahrtforachung, 1976, Bd.20, Nl, 36-38
12. Aviation Week & Rpace Technology, 1975, V.103, N.20, 53
13. Plug Revue Flugwelt Informationen, 1975, N21, 4-5
14.Aviation Week & Space Technology, 1974, V.101, Nl, 28-30
15. Aviation Week & Space Technology, 1973, V.99, x'10. 12-13
16. Aviation Space Environment, 1976, V.47, N4, 464
17. Aviation Week & Space Technology, 1976, V.104, N4, 49,61
18. Aviation Week & Space Technology, 1976, V.105, N1, 11
19. Aviation Week & Space Technology. 1976, V.104. N13, 53
20.AIM Paper. 1976, N7P3, 17.
21. Defesue/Space Business Daily, 1976, V.88, N34, 268,269
22. Aviation Week & Space Technology, 1976, V.104, N14, 52--36
11


Certain problems of space biotechnology

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