Rodent Habitat (RH) & Rodent Research (RR)
Concept of Operations
December 2013
ARC/Cecilia Wigley
Rodent Research Operations Manager
JSC/Brienne Shkedi
Research Integration Manager
JSC/Pete Hasbrook
Program Science Office
Contents
• Rodent Research Concept of Operations
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Project Overview
Hardware
Operations
Rodent Research – 1 Mission Overview (SpaceX-4)
Operations Requiring Two Crew
Procedures Suggested for RS Crew Member Participation
• RR-1 Data sharing opportunities
Project Overview
• Rodent Research (RR) includes of a suite
of hardware developed by ARC
– Rodent Habitat – provides on-orbit housing
for rodents in an EXPRESS rack
– Transporter – provides housing for rodents
during ascent on Dragon
– Animal Access Unit (AAU) – Interfaces with
both the Habitat and Transporter for
transfer of the animals between the units
and access to the animals for science
operations
– Hardware Support Kits (foodbars, clean up,
and containment bags)
– Experiment Unique Equipment (Specific to
each flight based on science requirements)
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Ex: Anesthesia Kit and Cardiac Puncture Kit
being developed for RR-2 (on SpX-6)
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Project Overview
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Other Projects Supporting RR Operations and Science
– Wetlab-2 (ARC)
• Provides modular, plug-and-play equipment that enables in situ sample collection,
processing and analysis for space biology and biomedical research
• Flying with RR-1 on SpX-4: Basic Animal Processing System
– Animal Health Kit, Anesthesia Kit, Basic Animal Dissection Kit
– Bone Densitometer (Techshot)
• Provides bone density scanning capability on-orbit
• Needed for RR-2 on SpX-6
– Wetlab Kit (JSC Biostronautics Contract)
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Provides various generic lab equipment for
sample processing and manipulation
– Rodent Tissue Box (CASIS/Bioserve)
• Provides formalin and ethanol fixation of hind limbs
• Needed for RR-1 and RR-2
– MSG Life Science Ancillary Hardware (LSAH) (MSFC)
• Enhancements to support biological handling and clean up
• Air Filtration System, Exchangeable Glove System, and
Decontamination System
• MSG Rear Wall Cover and Rodent Dissection Table also
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built by MSFC as a subset of this project.
Rodent Habitat Hardware Overview
• Based on existing AEM design
Rodent
Habitat
With new
Access Unit
mating access
lid
• Flown 27 times on Shuttle
• Modified to meet ISS needs
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Side A
Side B
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Rodent
Habitat
with lid
removed
November 14-15, 2012
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Reduce acoustics
Improve airflow
Accommodate floor lighting
Add cameras (4)
Change power and data (STELLA)
Single MLE unit houses 10 mice or 3 – 6 rats (2 on RR-1)
Front breather
Temperature and RH monitoring, no active thermal control
Transfer animals to a clean Rodent Habitat with a full
complement of food and water after 20 - 30 days to
achieve longer duration missions
Improved science and animal husbandry through video
monitoring and in-flight access
Adapted to operate at the ISS/EXPRESS Rack
Launches and returns as passive stowage
Powered only during duration of experiment operations
Animals housed in two groups of five on either side of the
Habitat
NASA Ames Research Center
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Transporter Overview
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Front Breather
Transporter
Rodents are launched in the Transporter
– Single MDL
– Current design is a front breather, but
potentially switching to a rear breather
– Power ~35 W for fans and lights
– Each unit houses up to 20 mice. Multiple
units may be flown depending on science
requirements.
– Animals loaded in the Transporter at L-25 hrs
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Or as late as possible.
Transporter certified for 10 days
– Would support up to 2 launch attempts before
change out with a new Transporter
Ducting used on
SpaceX for
airflow
Generic RR Concept of Operations
AEM
EXPRESS
Racks
(same or next) SpX
4. ISS Dock and
Animal Transfer
SpX
NLT Dock + 48 hours,
transfer animals from
Transporters to Rodent
Habitats (w/ AAU). Stow
Transporter for return.
Empty
AEM-T
6. Transfer
Remaining Live
Animals and
samples
5. On-Orbit
On-Orbit ConOps: Animal Processing System
Dissections and DXA using the
Animal Processing System and
Bone Densitometer
3. Ascent
~ 3 days
2. Launch
SpX
Mouse
Transfer
Box
Animal
Processing
System
(same or next) SpX
MSG
SpX
Stabilized
samples
Descent
Tissue
and
Blood
Samples
DXA
Cold
Stowage
Empty
AEM-T
Bioprocessing
and Analysis
System
Access
Unit
Habitat
7. Descent
Mouse
Transfer Box
8. Sample
Recovery
Activities may require MSG
1. Late Load
Transporters
Transporter
w/ animals
(20 mice, 10
days support)
Rodent Habitat
w/o animals
(passive
transport)
Rodent Habitat
w/ animals (10
mice, 30-45
days support)
Frozen
Samples
Refrigerated
Stabilized
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RR-1 Mission Design (SpaceX-4)
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20 mice launched in one Transporter, 10 per side
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Transferred to 2 Rodent Habitats on-orbit
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10 NASA Mice: 16-week-old C57Bl/6 female
10 CASIS Mice: Ten 16-week-old female mice (5 MuRF-1 knockout and 5 wild type, C57Bl/6
background
NASA RH: 5 per side
CASIS RH: Split on each side 5 knockout/5 wild type
Ops for CASIS Mice (SpX-4 Return)
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0g exposure for minimum 21 days, but as long as possible
Dissection of all 10 mice as follows, to occur over a max 3 day period
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Alternate wild type/knockout mice,
1st hindlimb dissected and frozen,
2nd hindlimb be dissected and formalin-fixed. Transfer to ethanol 48 +6 hours,
Liver and spleen dissected from 5 wild type mice. Preserve per NASA protocol below.
Remainder of each mouse carcass frozen
Frozen samples returned at -32C or colder
Ops for NASA Mice (SpX-5 Return)
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Housed in RH for 30 days (± TBD, pending biocompatibility test end Sept)
Dissect liver and spleen from 2 animals
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Fix spleens in RNA Later.
Freeze livers in a cryovial.
Remainder of each mouse carcass frozen
Remaining 8 animals euthanized and frozen for body weight measurements
Frozen samples are returned at -80C
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First health
check ASAP
after hatch
open
RR-1 Timeline & Ops Constraints
Sw dock 03/28
Calendar
4/17/2014
4/18
Sat
Sun
4/21 4/22 4/23 4/24 4/25
ATV dock
2
3
4
5
6
7
Day
L+2-6d
Dragon
Berth
0
1
AHC
AHC
Transfer
CB
T Stow
WLC
Sat
8
Sun 4/28 4/29
9
10
RR Setup
WLC
11
4/30 5/1 5/2
Progress Dock
12
13
14
WLC
Sat
Sun
5/5
5/6
5/7
15
16
17
18
19
16
17
WLC
WLC
Activites
NASA RH Days
Crewtime (hrs)
IBI
0
0.25
0.3
0.6
1
2
3
4.58
4
0.1
NLT 48 hr after berthing; based on 10 day
Transporter Lifetime
Requires unpacking of Habitat and associated kits,
configuring Habitat for ops in EXPRESS
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6
0.5
0.1
7
8
9
10
CASIS support activity
NASA validation support activity
Dragon docked phase
Shared support activity
NASA min 30
days in RH
Freeze/pack CASIS samples for
return
Sw dock 03/28
Calendar
5/8
5/9
Sat
Sun
5/12
5/13
Day
20
21
22
23
24
25
WLC
WLC
5/14 5/15 5/16
Wakata undock
26
27 28
WLC
CB
Dissect
Dissect
TT
TT
5/19
5/20
29
30
Dragon
Undock
31
32
CB
NASA RH Days
Crewtime (hrs)
18
19
20
21
0.25
6.3
6.8
1.5
IBI
X Stow
22
3
CB
23
24
0.3
0.1
25
26
27
0.1
28
14
15
0.1
AHC: Animal Health Check
Transfer: Transporter to Habitats
WLC: Water Level Check
T stow: Bag and stow Transporter
X Stow: Bag and stow Habitat
Dissect: Euthanize & Dissect
Euth: Euthanize remaining animals
TT: Tissue Transfer
CBI: Cold Block & Ice Brick Insert
CB: Crew Brief (15 min)
Sun
Activites
13
0.4
Sat
WLC
12
0.1
CASIS
dissection
w/in 3 days
max
CASIS min 21
days in 0 g
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WLC
Dissect
+Euth
5/21 5/22 5/23
33
34
35
Sat
36
TT
29
30
31
0.25
5.9
1.25
5/25
5/26
37
LSAH
Clean
38
X Stow
32
33
34
35
3
9
36
Operations Requiring Two Crew
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Certain Rodent Research procedures will require 2 crew members
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Most important during initial ops and dissections until prime crew member is
comfortable with the activity.
ALL procedures performed handling the animals are done with both hands
inside the AAU or MSG. Second crew member needed as an assistant.
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– Confirmed during operations validation test (OVT) in August
– Recommended by MDS crew during debrief
– Animal Transfer from Transporter to Habitat
• Holding the flashlight
• Record keeping of animal number, type, condition
• Potential Simultaneous Ops: second crew can also assist with prepping the Habitats while
the primary crew member is configuring the Transporter and AAU
– Dissection and Tissue Transfer Operations
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Reading lengthy/multi-page procedures
Pointing out key steps (such as matching proper labels for animals and their tissues)
Record keeping as needed
Retrieving any missing items as needed (i.e. primary needs more gloves or gauze or other
lab supplies)
• Potential Simultaneous Ops: second crew member could also do CASIS tissue transfers in
the glove bag (day 22/23 on the timeline in my charts) while the primary crew finishes
the CASIS dissections.
– Some experiment specific training may be required for second crew member
depending on the level of participation.
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Limited space inside the MSG with RR equipment
set up. No room to bring procedures inside.
Operator with hands inside
the MSG relied on helper to
read procedures and point
out key steps during the
long dissection operations.
Transfer of animals
from Transporter.
Required helper
holding flashlight and
recording
number/type of
animals pulled from
the Transporter.
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Procedures Suggested for RS Crew Member
Participation
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Procedure reader for all activities except the initial transfer operations
Time keeper for dissections
LSAH set-up in the MSG
All cold stowage activities
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5.
assumes they have received systems training on MELFI and Glacier
Coldbag configuration
MELFI inserts
Glacier inserts
Formalin to Ethanol tissue transfer
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assumes CASIS is providing OBT for this activity
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Cleaning and stowing the AAU after animal access operations
Stowing kits at the end of sessions
Water level assessments
Configuring and bagging the Transporter and Habitats for stowage and return in
Dragon
10. Final cleanup of MSG
Estimated RR-1 Russian crew time: 25-30 hours;
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See 39/40 OOS for additional details
RR-1 Data Sharing Opportunities
• RR-1/SpaceX-4 Data to be provided to Russian Scientists
– Data from 10 NASA mice can be provided to Russian scientists
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All mice will be euthanized
Up to two will be dissected on-orbit (removal of liver and spleens).
Cadavers and specimens will be returned on SpaceX-5
Data can be provided after NASA analysis of returned samples – via geneLAB?
Tissue sharing is not available on RR-1
– CASIS data is proprietary and will not be shared with Russian scientists
• Possibilities for broader collaboration on future NASA Rodent
Research missions – requires more discussion
– Tissue sharing
– Russian-defined rodent specimens
– Expanded crew time sharing