Organisms in Space
Greg Leonard and Darren Hughes
Mains Associates
1
History - Dogs in Space
A Russian stray dog named Laika was the first
biological specimen to orbit the Earth. She
flew aboard the Sputnik 2 spacecraft launched
on November 3, 1959.
Laika became an international
celebrity, with several countries,
from Romania to Mongolia,
celebrating the event with
commemorative stamps.
2
History - Primates in Space
Ham
Chimpanzee
January 31, 1961
Sam
Rhesus monkey
Two flights:
1959 & 1963
Enos
Chimpanzee
November 29, 1961
Two orbits
3
Timeline: Key Milestones (1)
‘Enos’
Mercury 5
1961
First
primate in
orbit
‘Laika’
Sputnik II
1957
First
organism in
orbit
‘Felix’
AG1
(France)
1963
John Glenn
Mercury 6
1961
First American
in orbit
Yuri Gagarin
Vostock I
1961
First human
in orbit
First cat
in orbit
Biosat II
1967
First seeds
germinated
in orbit
Biosat I
1966
Bion 1
1973
First of 11
unmanned
Russian
biological
research
capsules
‘Arabella’
Skylab 3
1973
Spacelab 3
1985
First reusable
animal
laboratory in
orbit
NASA/Mir
1995-98
Neurolab
1998
First dedicated
neurology
research
program in
orbit
Future milestones
(ISS and beyond):
• First mammal born in
space
First
bacteria
in orbit
First
student
experiment
in orbit
First seedto-seed
growth of
plants in
orbit
• First biology
experiments beyond
Earth orbit
• First multi-generational
mammalian studies in
space
• First self-sustaining
ecosystem in space
4
Timeline: Key Milestones (2)
‘Enos’
Mercury 5
1961
First
primate in
orbit
‘Laika’
Sputnik II
1957
First
organism in
orbit
‘Felix’
AG1
(France)
1963
John Glenn
Mercury 6
1961
First American
in orbit
Yuri Gagarin
Vostock I
1961
First human
in orbit
First cat
in orbit
Biosat II
1967
First seeds
germinated
in orbit
Biosat I
1966
Bion 1
1973
First of 11
unmanned
Russian
biological
research
capsules
‘Arabella’
Skylab 3
1973
Spacelab 3
1985
First reusable
animal
laboratory in
orbit
NASA/Mir
1995-98
Neurolab
1998
First dedicated
neurology
research
program in
orbit
Future milestones
(ISS and beyond):
• First mammal born in
space
First
bacteria
in orbit
First
student
experiment
in orbit
First seedto-seed
growth of
plants in
orbit
• First biology
experiments beyond
Earth orbit
• First multi-generational
mammalian studies in
space
• First self-sustaining
ecosystem in space
5
Some Organisms Studied in Space
Bacteria
Aeromonas proteolytica
Bacillus mycoides
Bacillus subtilis
Bacillus thuringiensis
Burkholderia cepacia
Chaetomium globosum
Deinococcus radiodurans
Escherichia coli
Nematospiroides dubius
Rhodotorula rubra
Salmonella typhimurium
Trichophyton terrestre
Invertebrates
Acheta domesticus (Cricket)
Araneus diadematus (Spider)
Biomphalaria glabrata (Snail)
Caenorhabditis elegans (Nematode)
Cynops pyrrhogaster (Newt)
Drosophila melanogaster (Fruit fly)
Habrobracon juglandis (Wasp)
Manduca sexta (Tobacco hornworm)
Pelomyxa carolinensis (Amoeba)
Pothetria dispar (Gypsy moth)
Tribolium confusum (Beetle)
Trigonoscelis gigas (Beetle)
Plants
Aesculus hippocastanum L. (Horse chestnut)
Arabidopsis thaliana (Thale cress)
Avena sativa (Oat)
Brassica rapa (Field mustard)
Capsicum annuum (Ornamental pepper)
Ceratodon (Moss)
Ceratopteris (Fern)
Ceratophyllum demersum (Hornweed)
Cucumis sativus (Cucumber)
Dactylis glomerata L. (Orchard grass)
Daucus carota (Carrot)
Digitalis lanata (Foxglove)
Digitalis purpurea L. (Foxglove)
Elodea (Waterweed)
Flammulina velutipes, Agaricales (Fungus)
Glycine max (Soybean)
Haplopappus gracilis (Haplopappus)
Helianthus annuus L. (Sunflower)
Hemerocallis (Daylily)
Lepidium sativum (Garden cress)
Linum usitatissimum (Flax)
Lycoperscion esculentum (Tomato)
Neurospora crassa (Fungus)
Nicotiana tabacum (Tobacco)
Oryza sativa (Rice)
Physarum polycephalum (Slime mold)
Pseudotsuga menziesii (Douglas fir)
Pseudotsuga taeda (Loblolly pine)
Saccharomyces cerevisiae (Yeast)
Tradescantia (Spiderwort)
Triticum aestivum (Wheat)
Triticum vulgare (Wheat)
Vigna radiata (Mung bean)
Zea mays (Corn)
Vertebrates
Canis familiaris (Dog)
Felix maniculata (Cat)
Homo sapiens (Human)
Macaca mulatta (Rhesus monkey)
Macaca nemestrina (Pigtail macaque monkey)
Mus musculus (Mouse)
Oryctolagus cuniculus (Rabbit)
Pan troglodytes (Chimpanzee)
Perognathus longimembris (Pocket mouse)
Rattus norvegicus (Rat)
Saimiri sciureus (Squirrel monkey)
Testudo horsfieldi Gray (Tortoise)
Birds
Coturnix coturnix (Quail)
Gallus gallus (Chicken)
Aquatic species
Arbacia punctulata (Sea urchin)
Aurelia aurita (Jellyfish)
Fundulus heteroclitus (Killifish)
Lytechinus pictus (Sea urchin)
Opsanus tau (Toadfish)
Oreochromis mossambicus (Cichlid fish)
Oryzias latipes (Medaka fish)
Rana catesbeiana (Bullfrog)
Rana pipiens (Frog)
Strongelocentrotus pupuratus (Sea urchin)
Xenopus laevis (Frog)
Xenopus laevis Daudin (South African toad)
Xiphophorus helleri (Swordtail fish)
6
Benefits of Studying Different Organisms
Benefits to Space Exploration
• Risk mitigation
• Medical care
• Life support
Benefits to Life on Earth
• Biology
• Medicine
• Technology
• Education
7
Human Studies in Space
• Bone Deterioration
• Muscle Atrophy
• Cardiovascular Deconditioning
• Immune Suppression
• Sleep Disturbances
• Balance Disorders
8
Why Not Just Study Humans in Space?
Primary Reasons:
• Ethical
• Practical
• Biological
• Medical
• Logistical
9
Why Study Microbes in Space?
• Basic research
• Bioregenerative
life support
• Nanotechnology
10
Why Grow Plants in Space?
• Basic research
• Food source
• Remove CO2
• Produce O2 & water
vapor
• Psychological benefits
11
Current and Future Directions
• Focus on cell and molecular biology
• Understanding of underlying mechanisms
• Use of “model” organisms
• Reference studies
12
Model Organisms
• Well-characterized
• Genetically sequenced
• Appropriate for space research
13
Escherichia coli (Bacteria)
Bacteria (E. coli) 0.5-1.5µm
Growth Requirements
Microfluidics
Liquid culture likely for flight, can be grown on solid medium
Sensors
pH, oxygen, carbon dioxide,temperature
Temperature
Will grow between 10 - 45°C; Optimum 37°C
Salinity
Tolerates low to moderate salinity
Nutrients
LB for bacteria: yeast extract, bacto-peptone, sodium chloride, water
pH
Growth optimum between pH 5.5 - 8.0
Doubling rate
20 minutes to several hours
Light
Not required
Aeration
E. coli is facultative anaerobe, i.e. does not require O2 but grows better in its presence
Wastes
Gaseous (CO2) and liquid (metabolites)
14
Saccharomyces cerevisiae (Yeast)
Yeast (S. cer.) 5-12µm
Growth Requirements
Microfluidics
Liquid culture likely for flight, can be grown on solid medium
Sensors
pH, oxygen, carbon dioxide,temperature, pressure
Temperature
Will grow at temperatures between 3 - 40°C; Optimum 28°C
Salinity
Yeast grows within a wide range of salt concentration
Nutrients
YPD : yeast extract, bacto-peptone, glucose, water
pH
Growth optimum between 5.0 - 6.5
Doubling rate
1-4 hours
Light
Not required
Aeration
Does not require O2 for growth, but grows better in its presence
Wastes
Gaseous (CO2) and liquid (metabolites)
15
Caenorhabditis elegans
(Nematode)
Growth Requirements
Microfluidics
Sensors
Temperature
Salinity
Nutrients
pH
Doubling rate
Light
Aeration
Wastes
Liquid culture or solid culture. Organism is ~ 1mm in length and grows in axenic
defined liquid media or on solid media using bacteria as food
carbon dioxide, oxygen, temperature
17°C optimum for growth; heat shock at 25°C; 30°C for > 20 hrs is lethal
0.1 - 0.5 molar simple inorganic salts (NaCl, KHPO4), wide tolerance range
Consumes dissolved nutrients in axenic liquid culture media, or on solid media
consumes bacteria (E. coli).
Optimum pH 6.0; tolerates pH 3 - 9
3-6 days to mature; temperature dependent
Generally not required
Chamber ventilation required for axenic liquid culture in gas-permeable opticell
cartridges or for culture on solid media
Gaseous (CO2), liquid (metabolites), and debris (dead worms)
16
Drosophila melanogaster
(Fruit fly)
Growth Requirements
Microfluidics
Sensors
Temperature
Nutrients
pH
Doubling rate
Light
Aeration
Wastes
Solid medium
pH, oxygen, carbon dioxide,temperature
22 - 30°C with heat shock at 45°C.
YPD : yeast extract, bacto-peptone, glucose, water
~ 6.5 - 5.0 optimum; range ~7.2 – 4.5
1-4 hours
Not required
Active or passive aeration required
Gaseous (CO2) and liquid (metabolites)
17
Arabidopsis thaliana (Brassica)
Growth Requirements
Microfluidics
Sensors
Temperature
Humidity
Nutrients/water
Doubling rate
Light
Gas Composition
Wastes
Water/nutrient delivery system
pH, carbon dioxide, oxygen, ethylene, temperature, light
17 - 25°C; some protocols call for 15°C during dark cycle
65 - 100%; vegetative phase tolerant of a broad relative
humidity range but above 855 can affect flowering and seed set
Consistent with soil composition; well aerated soil required
4-8 week growth cycle
16hr light, 8 hr dark cycle; light intensity 250 u mol
Typical air composition: 21% O2, 78% N, 0.05% CO2
O2, ethylene
18
Mammalian Cells
Growth Requirements
Microfluidics
Sensors
Temperature
Humidity
Salinity
Nutrients
pH
Doubling rate
Light
Aeration
Wastes
Liquid culture flowing above an adherent cell layer or cells growing in suspension in a
liquid culture
pH, CO2 , O2 , temperature, pressure, flow rate
Tolerate only a very narrow temperature range, typically between 37°C - 42°C. Optimal
temperature is 37°C ± 0.5°C
~ 80% in incubators
A variety of commercial buffered saline solutions used; most popular are Hank’s and Earle’s
Different cell types require different media formulations, e.g. DMEM: glucose, Lglutamine, sodium pyruvate, phenol red. Between 5-20% fetal bovine serum or horse
serum is a common supplement.
pH range between 6.8 - 7.2
12 - 48 hours
Not required
Air, as oxygen source, must be added to culture in a way to avoid sheer stress to which
mammalian cells are very sensitive. CO2 levels must be kept at certain level; typically 5%
19
Gaseous (CO2) and liquid (metabolites)
Rodents
Housing & Husbandry Requirements
Habitat
Rodent cage: ventilated and kept free of contaminants from urine & feces
Sensors
O2 , CO2 , temperature, activity (video)
Temperature
18°C - 26°C
Humidity
30 - 70%
Food/WaterIrradiated food bars (rodent chow) with long shelf-life / automatic watering
manifolds or water bottles
Population Density
6 rats or 10 mice per cage
Light
8-10 hours/day exposure during circadian cycle
Ventilation
Control O2 , CO2 , particulate contaminants, animal odors
Wastes
Urine, feces must be contained
20
Ethical Use of Animals at NASA
Bioethical Principles
• Respect for life
• Societal benefit
• Nonmalificence
21
Regulations and Oversight
• IACUC and federal regulations
• Scientific standards
• Agency oversight
• Public scrutiny
22
References
Medical Operations of Space Flight I
Dr Arthur Arnold, Jr
Kennedy Space Center
Life into Space
Space Life Sciences Experiments 1991-1998
Eds. K Souza, G Etheridge & P. X. Callahan
Model Organisms for Space Biology Research
Dr Rita Briggs
Lockheed Martin
Fundamentals of Space Biology
Eds M. Asashima & G.M. Malacinski (1990)
Japan Scientific Societies Press & Springer-Verlag
Early History of Space Biology and Medicine
John P. Marbarger
Acta Astronautica Vol. 43 No. 1-2 pp 9-12 1998
International Flight Experiments Database
http://www.mainsgate.com/IFE/index.html
NASA Life Sciences Data Archive
http://www.lsda.jsc.nasa.gov
23