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3 ESA experiment to be performed in Biolab The scientific aim is to study how mammalian cell cultures in vitro react specifically to microgravity Two different cell lines, both of human origin, will be tested; a fibroblast cell line (synthesizes the extracellular matrix and collagen) and an osteoblast cell line (synthesizes bone) Computer-assisted image analysis of PFA-fixed human skin fibroblasts Background
Citation preview
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POIWG #38, July 2015
BIOTESC USOC, Switzerland
Martin Haag & Jeannine Winkler
BIOTESC e-mail:[email protected]
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3
ESA experiment to be performed in Biolab
The scientific aim is to study how mammalian cell cultures in vitro react specifically to microgravity
Two different cell lines, both of human origin, will be tested; a fibroblast cell line (synthesizes the extracellular matrix and collagen) and an osteoblast cell line (synthesizes bone)
Computer-assisted image analysis of PFA-fixed human skin fibroblasts
Background
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Hardware:
6 different Integrated Advanced Experiment Containers (EC) are uploaded at ambient temperatures. Three EC are to be fixated with RNAlater and three with PFA.
EC
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Hardware: UploadFixative Reservoir Single
Sample Reservoir
EC
Parts of the hardware are uploaded already mounted inside the ECs
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Hardware: UploadFixative Reservoir Single
Sample Reservoir
Med Reservoir/Fixative Reservoir Double
Culture Chamber
Type 1 EC
EC
The other parts of the hardware are uploaded at -80°C inside Type 1 ECs and need to be integrated into the EC on orbit
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Hardware: Assembly
Culture Chamber
EC
The assembly of one EC is expected to take up to 1 hour to complete
Fixative Reservoir Single
Sample Reservoir
Med Reservoir/Fixative Reservoir Double
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Hardware: Disassembly
Culture Chamber
EC
Only the ECs fixed with RNAlater are disassembled and downloaded at -80°C inside Type 1 ECs. The ECs fixed with PFA are downloaded in one piece at +4°C
Sample Reservoir
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Experiment Overview
SpXdocking
Transfer and storage on ISS
Experiment start
zero time control run
0g run
1g run
Recovery period:
37°C, 1 g38 ± 2 h
Fixation/Medium Replacement
37°C, 1 g38 ± 2 h
37°C, 1 g38 ± 2 h
Disclaimer:Only two ECs fit on the Biolab Rotor at the same time and the experiment will be run sequentially!
Three runs!
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Experiment Overview
SpXdocking
Transfer and storage on ISS
Experiment start
zero time control run
0g run
1g run
Recovery period:
37°C, 1 g38 ± 2 h
Fixation/Medium Replacement Fixation
Active phase:
37°C, 1 g38 ± 2 h
37°C, 1 g38 ± 2 h
37°C, 0 g34 ± 1 h
37°C, 1 g34 ± 1 h
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Key points• Upload with SpX:
• -80°C: 56 Culture Chambers (in 28 Type 1 EC), 10 Med Reservoirs (in 10 Type 1 EC)• Ambient: 1 Zero EC PFA, 1 Zero EC RNA, 2 Nominal EC PFA, 2 Nominal EC RNA
• After Docking: Transfer to cold stowage (MELFI -80°C and BIOLAB TCU -20°C) • 6 Experiment Containers, processed sequentially in 3 runs: 11 days of
experiment!• Download with SpX:
• -80°C: 2 Sample Reservoirs (in 2 Type 1 EC)• -20°C: 28 Culture Chambers (in 14 Type 1 EC)• +4°C: 1 Zero EC PFA, 2 Nominal EC PFA (all PFA ECs are downloaded fully assembled)
Responsibilities
• NASA: upload/download, transfer to COLUMBUS/SpX, Transfer to MELFI• MUSC: for all ground activities, Biolab facility related crew questions• BIOTESC: experiment related activities, scientific questions
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Challenges
• Very demanding timeline, few break points to pause • Once the cells are thawed extremely strict margins to fulfill • Planning: days with dense timetable for crew
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SPHEROIDS – Experiment Overview
Scientific Aims:• comparing the effects of real microgravity to those
induced by mimicked weightlessness on the RPM • evaluating the protective effect of VEGF against potential apoptosis• post-flight analysis of three dimensional cell assembly,
differentiation and apoptosis
H/W upload:
SpX-7: KUBIK Interface Plates (KIPs) (lost)
SpX-8: SPHEROIDS Experiment Containers
2x
8x
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Impact of SpX-7 Loss and Spheroids Flight Readiness
• 2 KIPs can be replaced by 1 spare FM and upgrade of 1 QM (upgrade feasible in 3-5 months)
• 2 KIPs need to be uploaded before the Experiment H/W
• KUBIK 6 is at the moment not operational due to expired batteriesin centrifuge eBox
• 2 KUBIK eBoxes were lost on SpX-7
• Refurbished eBoxes from KUBIK 3 will have to replace the lost onesand be integrated in KUBIK 6 before the experiment
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KUBIK Overview
D1 D2
• KUBIK Interface Plates (KIPs) – installed at least 1 day before experiment start
• KUBIK 5 – 39 hours incubation at 37˚C => MELFI cold stowage
• KUBIK 6 – 8.5 days incubation at 37˚C => change of temperature to 6˚C and cold stowage in KUBIK up to 30 days
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Experiment Activities- Day 0Big picture wordsKUBIK 5 setup & KIP installation, KUBIK 6 setup & KIP instalation
- Day 1Experiment installation in KUBIK 5 and KUBIK 6
- Day 3Experiment removal from KUBIK 5 & transfer to MELFI
- Day 3 – 9KUBIK 5 datacopyRegular checks of KUBIK 6 temperature (37˚C )
- Day 10KUBIK 6 datacopySetting KUBIK 6 to 6˚C
- Day 10 - downloadRegular checks of KUBIK 6 temperature (6˚C )
- Download dayExperiment removal from KUBIK 6 and MELFI and packing
- 1 day after downloadKUBIK 6 datacopy 2
SPHEROIDS-KUB5-SU
SPHEROIDS-KUB6-SU
KUB5-KIP-INST
KUB6-KIP-INST
SPHEROIDS-KUB-PREP
SPHEROIDS-EC-INST
SPHEROIDS-KUB5-DEINST
SPHEROIDS-KUB5-DATACOPY
SPHEROIDS-KUB6-DATACOPY
INC
UB
ATIO
N a
t 37
˚C 3
9 hr
s
INC
UB
ATIO
N a
t 37˚
C
207
hrs
SPHEROIDS-COOLTEMP
SPHEROIDS-KUB6-DEINST
SPHEROIDS-EC-PACK
SPHEROIDS-KUB6-DATACOPY 2
Challenges / Watch Outsi. Time constraints are given by the automatic,
pre-programmed timeline in the EC:i. Removal of EC before end of experiment run will lead to complete loss of science (no fixation)
ii. In case of KUBIK power loss the timeline is interrupted by the duration of the power loss –> de-installation activity (for ECs in KUBIK 5) and/or setting of KUBIK to 6°C activity (ECs in KUBIK 6) to be shifted in the timeline accordingly
iii. The experiment run between EC installation and removal/cold stowage activity is 39hrs/207hrs: EC installation and EC removal/cold stowage activity need to be coordinated such that they fit in the crew day
ii. In case of KUBIK anomaly the recovery action shall be started as soon as possible: gradual loss of sample viability and complete loss of science after 12 hours
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Objectives of EPO Peake
• To strengthen and to encourage the teaching of science and thus stimulate the curiosity of students and motivate them towards further study of STEM (Science, Technology, Engineering, and Mathematics) and computing and coding subjects
• To primarily make the students aware that conditions on the ISS are different to those on earth and that in weightlessness, effects can be detected that are very hard to detect on Earth.
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Overview of EPO Peake
• Educational Program Operations of Timothy Peake• Joint project between UK Space Agency (UKSA) and ESA• Four different projects
Seeds in Space o One activity: selfies of Timothy Peake with seeds (during day pass)o Post flight growth difference analysis of seeds that were in space and on
ground done by studentso Location: Cupolao Inc46 crew time: 30min prime o Task List or hard scheduled
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Overview of EPO Peake
Classroom Videos Demonstrations (CVD)o One activityo Five demonstrations on physical laws
(describing motions and interactions of physical entities on orbit)o Location: Columbuso Inc46 crew time: 2hrs reserve o Task List
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Overview of EPO Peake
Astro Pi o Credit card sized Raspberry Pi computer which will execute codes
programmed by studentso Two Astro Pis will be uploaded: one with a standard and one with an
infrared camerao Insertion of SD card in Astro Pi to start the computer and execute the
students codeso Download of output files from Astro Pi activities stored on SD card,
distribution to schoolso Locations: Columbus, Cupola, ev. US LAB (three deployments required)o Upload: 45So Inc46 crew time: 50 min o Hard scheduled prime and reserve
More details in splinter, July 23rd at 13:00
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EPO-ASTRO-READ (00:10)
EPO PEAKE AstroPi
CREW
EPO-ASTRO-DEINST(00:15)
EPO-ASTRO-STOW(00:05)
EPO-ASTRO-UNSTOW
(00:20)
EPO-ASTRO-COL-SU (00:25)
Code execution
EPO-ASTRO-FILECOPY
(00:20)
EPO-ASTRO-FILECOPY
(00:20)
Astro Pi FlowChart
Execution of the same sequence for each deployment
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Overview of EPO Peake
Generic Videoso 13 videos (script recording) o To provide scientific information, the astronaut’s views on earth
or linking experiment activities via visual and audio mediao Locations: Cupola, Columbus, US LAB, JEMo Inc46 crew time: 1hr 10 min prime, 6hrs reserveo Task List (gap fillers)
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Challenges
• Loss of CVD Kit and Seeds with SpX-7
• Re-launch possibilities: - 44S (Prime path), 61P (21.09.2015), Orb4
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• Located in Hergiswil, Switzerland
• KUBIK Facility Responsible Center (KUBIK in Columbus since Inc. 23)
• BIOLAB Facility Support Center
• Responsible USOC for ESA Thermal Container, EPO Experiments, NightPod
BIOTESCBiotechnology Space Support Center
