NASA Bioculture System Capabilities
•
Provides 10 individual cassettes containing hollow fiber bioreactors with media
perfusion for a no shear cell culture environment for both adherent and non-adherent
cell types, including 3-D tissues and microorganisms
•
Supports automated and manual inflight inoculations for growth initiation,
stimulation, sub-culturing, infection, and fixation; also supports automated and
manual sample removal for inflight microscopy, analysis and/or fixation
•
Provides programmable temperature settings, +4-39oC
•
Includes fluid loops and a gas exchanger for CO2 and O2 perfusion
•
Supports design flexibility to meet investigator culture chamber, fluid circulation, and
automated and manual specimen processing requirements including maintaining
cultures over long durations
•
Downlinks individual cassette data; incubation temperature, cold chamber
temperature, CO2, humidity, power, pump activity and event activation
•
The following are examples of cells flown in the Cell Culture Module (precursor to the
Bioculture System) on the Shuttle:
Skeletal myoblasts
Human osteoblasts
Human lung fibroblasts
Stem cells (hematapoetic)
Rat glial tumor cells
Elastin heterografts
Human osteoblast-like cellsChicken Osteoblasts
Primary hamster fibroblasts
Human colon carcinoma cells
Chick embryo osteoblasts
Bovine aortic endothelial cells
Rat skeletal muscle cells
Rat osteoblasts Adipose derived
Mesenchymal cells (cartilage) mesenchymal stem cells
Bioculture System (baseline design)
Gas Supply Assembly
Cassette
Cover
Power
Supply
Module
Cassette Assembly
Media, sample
collection,
Injection fluid, and
sump bags
GoreVents
Incubation
Chamber
Fan Plenum
Insulation
Cassette
Front Plate
Cold
Chamber
Cassette
Base
Biochamber
Oxygenator
Inject
Solution
Pump
Media
Bag
Sample
Collection
Fluid Flow
Path Schematic
Solenoid
Manifolds and Valves
Biochamber
Media
Waste
Bag
Heating
Loop
Incubation Chamber
Cold Chamber
16 solenoid valves
for up to a total of 16
sample/injection bags
Bioculture System Specification
Nominal Configuration
Specimens: Cell culture, tissue culture, 3D cultures,
explant cultures, and microbes
•
•
•
Docking Station
– Power Supply Module
– Gas Supply Assembly
– Cassette power and gas connectors
– ExPRESS Rack mounted
10 Cassette Assemblies in the Docking Station
• Independent fluidics and environmental
control except for the gas supply
Each Cassette Assembly contains:
 1 Biochamber
 Flow Path Tubing
 Pump
 Up to 16 Solenoid Valves
 Up to 16 sample/injection bags
 ~200 ml Media Bag
 1 Sump waste bag
 Memory card with set point
programming for operational parameters
 Cold Chamber
 Incubation Chamber
 Oxygenator
 Heating Loop
 Temperature and Humidity Sensors
 Three levels of containment
o 2 levels provided by the cover and
1 level provided by the flow path
Bioculture System Specification for Experiments
Environmental Control per Cassette Assembly
• Incubation Chamber Set Points: Ambient to +42 oC
• Cold Chamber Set Points: Ambient to +5 oC
• Temperature is experiment defined and independent for
each Cassette Assembly
Gas Supply – Shared between all Cassette Assemblies
• Nominal 5% CO2, 20% O2, balance Nitrogen
• Investigator defined gas composition
• Shared between all Cassette Assemblies
Fluid System per Cassette Assembly
• Pulsatile, intermittent, and continuous modes
• Pump: 0 – 15 ml/ min
• Micro-solenoid diverter valves
• Biomedical grade tubing
• Replaceable Media bag and separate waste sump bag
• 16 replaceable sample/injection bags
• Programmable delivery schedule or continuous recirculation
for medium
• Programmable schedule for sampling and injections
• Replaceable Biochamber
• System compatible with with most fixatives and
preservatives
• Membrane oxygenator using silicone elastomer tubing
• Heating loop to pre-warm the medium
Total of 3 levels of containment
The fluid system can be accessed by the Crew on ISS
Cassette Assemblies fit in the ISS Microgravity Sciences
Glovebox
Examples of Potential ISS Supplemental Equipment for
Cell, Tissue, and Microbiology Research
• ISS Science facilities/equipment:
http://www.nasa.gov/mission_pages/station/research/facilities_category/index.html#.Ugqdn
xY9_Yk
• Microgravity Glove Box
• Light Microscopy Module for microscopic analysis
• Wetlab-2 RNA SmartCycler qRT-PCR System for real time gene
expression data downlinked to investigator
• Small centrifuge for 1-g, fractional-g (e.g., European Modular
Cultivation System [EMCS], Kubik), or sample processing (e.g.,
Cell Biology Experiment Facility [CBEF], Biolab)
• Refrigerators and freezers
Science program reference documents
• National Research Council’s 2011 Decadal Survey
Report, “Recapturing a Future for Space Exploration:
Life and Physical Sciences Research for a New Era”
(http:www.nap.edu/catalog/13048.html)
• NASA Fundamental Space Biology Science Plan 20102020
(http://www.nasa.gov/sites/default/files/files/10517_F
SBPlan-Signed_508.pdf)
Check one: Space Bio__X__, HRP___, DoD___, CASIS/Commercial___
Assumptions:
1) Bioculture System Docking Station housing 10 Cassettes per flight
2) Repeat experiments during flight are possible provided additional cassettes are available
3) Bioculture System docking station remains on-orbit or returns carrying the Cassettes
4) Nominal mission duration: Bioculture System, up to 60 days
Type of Biochamber
Culture Initiation
Solution
Injection
Other or
Real-Time Method:
Custom
microscopy; Automate
(Specify
On-Orbit video/photo and/or
Type[s]) Pre-flight On-orbit Sub-culture
(Y/N)
Manual
Flatbed
culture
Murine
surface
Y
60
10
2
20
Y
N
2
Y
Y
Y
Automated
Stem Cell
with
(white light)
hollow
fibers
1)Obtain data to investigate stem cell proliferation in microgravity; 2) Investigate if the potential for stem cell differentiation induction changes with prolonged
exposure to microgravity
Total
Quantity of
Science/Commercial Theme per Experiment
Number of
quantity
Live
Hollow Biochambers
Experiment No. ________
Experim't
replicates Experim't
of
Specimen
Fiber
housed in a
Duration; Type of
per
runs per Cassettes return Bioreactor
single
Days
Specimen specimen
flight
required
(Y/N)
(Y/N)
Cassette
Theme: Stem Cell Proliferation and
Differentiation
a. Experiment Objective(s)
b. Cell culture system of choice
NASA Bioculture System
c. Special equipment required for on-orbit
operations (manual and automated)
d. Specimen removal for on-orbit analysis?
Type of analyses or sample isolation?
e. Special considerations for Crew training
and inflight Operations?
f. NASA relevance linkage: e.g., NRC
Decadal Survey &/or Space Biology Plan,
2010; Non-NASA relevance linkage
g. Does this experiment require multiple
flights to accomplish the stated objectives?
How many flights?
h. Does the specimen require a specific gas
environment? What % composition
i. Types of solution for injection (additives,
drugs, staining solutions, etc) ?; multiple
injection time course? Identify Tox level if
known
j. Specimen sampling ? Multiple sampling
time course?
k. Co-culturing needed (e.g. on-orbit
infection, cell to cell interactions, etc)
1) Culture initiation injection tool, 2) warming tool - the cells for initiating cultures on-orbit are flown as frozen in vials
l. Additional sensors desired
pH
m. "omics" analysis planned?
RNA, protein, metabolic products
Yes - on-orbit isolation of RNA and DNA
Method of proper thawing of cells
Decadal Survey ; Space Biology Science Plan
Requires 2 flights - 1st flight identifies target differentiation pathways; 2nd flight provides experiments for indepth analyses
No, 5% CO2 with standard atmospheric O2 and N2 as balance
1) Paraformaldehyde, 2) RNALater, 3) Trizol, 4) antibodies for differentiation markers, and 5) compound solutions for inducing differentiation
Yes, 1 x before induction of differentiation
No
Specimen
Sampling
Method:
Automate
and/or
Manual
Manual
Any noteworthy assumptions?
Check one: Space Bio___, HRP___, DoD___, CASIS/Commercial___
1
2
3
Type of Biochamber
Culture Initiation
Solution Specimen
Total
Quantity of
Injection Sampling
Science/Commercial Theme per Experiment
Number of
quantity
Live
Hollow Biochambers Other or
Real-Time Method: Method:
Experiment No. ________
Experim't
replicates Experim't
of
Specimen
Fiber
housed in a Custom
microscopy; Automate Automate
Duration; Type of
per
runs per Cassettes return Bioreactor
single
(Specify
On-Orbit video/photo and/or
and/or
Days Specimen specimen
flight
required
(Y/N)
(Y/N)
Cassette
Type[s]) Pre-flight On-orbit Sub-culture
(Y/N)
Manual
Manual
Theme:
a. Experiment Objective(s)
b. Cell Culture System of Choice:
c. Special equipment required for on-orbit
operations (manual and automated)
d. Specimen removal for on-orbit analysis?
Type of analyses or sample isolation?
e. Special considerations for Crew training
and inflight Operations?
f. NASA relevance linkage: e.g., NRC
Decadal Survey &/or Space Biology Plan,
2010; Non-NASA relevance linkage (Provide
specific quote/priority number, or other
identify designator.
g. Does this experiment require multiple
flights to accomplish the stated objectives?
How many flights?
h. Does the specimen require a specific gas
environment? What % composition
i. Types of solution for injection (additives,
drugs, staining solutions, etc) ?; multiple
injection time course? Identify Tox level if
known
j. Specimen sampling ? Multiple sampling
time course?
k. Co-culturing needed (e.g. on-orbit
infection, cell to cell interactions, etc)
l. Additional sensors desired
m. "omics" analysis planned? If yes, specify